Mechanistic insight into inhibition of two-component system signaling

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1 Supporting Information Mechanistic insight into inhibition of two-component system signaling Samson Francis, a Kaelyn E. Wilke, a Douglas E. Brown a and Erin E. Carlson a,b* a Department of Chemistry, Indiana University, 800 E. Kirkwood Avenue, Bloomington, Indiana, USA. b Department of Molecular and Cellular Biochemistry, Indiana University, 212 S. Hawthorne Drive, Bloomington, Indiana, USA. Corresponding Author: carlsone@indiana.edu Contents Supporting Information Figures and Tables..2 Docking-Based Virtual Screening General Docking of Compounds In Vitro Activity Assays General Methods and Information Experimental Methods HK853 Construct (Thermotoga maritima) PCR Site-Directed Mutagenesis for Generation of HK853 D411A Construct Circular Dichroism (CD) Spectroscopy of HK853 Proteins Rescue ABPP analysis of NH125 (3), 48, 49, 50 and Saturation Transfer Differential NMR Experiments Determination of K d Using STD-NMR Measurement of STD Amplification Factors References S1

2 Supporting Information Figures and Tables S2

3 Table S1. Rank and score of compounds reported to demonstrate TCS inhibitory activities and their corresponding Surflex score, rank, and documented biological data. # Structure Receptor 1ID0 Receptor 1I58 Receptor 2C2A Receptor 3CGY 1 Surflex Score Rank Surflex Score Rank Surflex Score Rank Surflex Score Rank TCS Tested IC 50 HK (µm) IC 50 RR (µm) MIC AlgR Ref AlgR AlgR1 AlgR2 CheA NR II KinA AlgR1 AlgR2 CheA NR II KinA HpKA-DrrA HpKA-DrrA S3

4 HpKA-DrrA VicK VicK VicK VicK > VicK > VicK YycG YycG S4

5 YycG YycG YycG YycG > YycG HpKA- DrrA YycG S5

6 KinA- Spo0F , KinA- Spo0F KinA- Spo0F KinA- Spo0F NR II - NR I KinA- Spo0F KinA- Spo0F KinA- Spo0F > KinA- Spo0F , 10 S6

7 KinA- NR II KinA- NR II KinA- NR II KinA- NR II KinA- NR II NR I- NR II KinA- Spo0F 2.2 KinA- Spo0F NR I- NR II HprK/P NR II S7

8 YycF- YycG YycF- YycG SF- EnvZc SF- EnvZc SF- EnvZc YycF- YycG YycF- YycG S8

9 Fig. S1 The fragment-guided docking approach in Surflex. A portion of the co-crystallized ligand (shown with the adenine moiety in the receptor PhoQ, PDB: 1ID0) is retained and utilized to guide the placement of compounds in the active site. Fig. S2 The docked conformation of AMP-PNP in the active site of PhoQ superimposed with the adenine moiety (orange) of the co-crystallized structure. The computed RMSD between these two moieties was S9

10 Fig. S3 Predicted binding poses of selected TCS inhibitors in the nucleotide-binding region of HKs. Some portions of the protein have been cut away for clarity. (A) Compound 4 interacting with Asp449 in receptor 1I58. (B) The interaction of compound 15 with Asp411 in receptor 2C2A. (C) Asp415 interacting with inhibitor 20 in receptor 1ID0. (D) Salt-bridge interaction formed between inhibitor 30 and Asp416 of 3CGY. Fig. S4 Cluster of guanidine-bearing compounds (26, 27, 30, 33) and their corresponding interactions with Asp416 of PhoQ (3CGY; E. coli). S10

11 H Box Input_pdb_SEQRES_2C2A M E N V T E S K E L E R L K R I D R M K T E F I A N I S H E L R T P L T A UniRef90_A5IIT0_236_ V E N V T E S K E L E R L K R I D R M K T E F I A N I S H E L R T P L T A UniRef90_B9KAB5_220_ V E N I T A S K E L E R L K R I D R M K T E F V A N I S H E L R T P L T A UniRef90_UPI000210F96E_244_ D V T N T K E L E R L K Q I D A L K T E F I A N V S H E L R T P L A A UniRef90_B7IDY9_224_ N V T Y E I E L E K L K Q L D K M K T E F V A N I S H E L R T P L T A UniRef90_A6LN40_223_ N V T Y E M E L E K L K Q L D K I K T E F V A N I S H E L R T P L S A UniRef90_A8F6Z9_244_ D V T N T K E L E K L R A A D R L K T D F I A T V S H E L R T P L T A UniRef90_A7HK36_224_ I R N I T S E K E V Q K L K E L D K I K T N F I A N I S H E L R T P L A A UniRef90_D5WTD2_380_ I H D V T E R R R L E Q V R K E F V T N V S H E L K T P V T A UniRef90_Q67RF9_205_ R D I T R S R Q L E Q M R T E F V A N V T H E L R T P L T S UniRef90_B3E3A4_350_ L K R V E T M R R D F V A N V S H E L R T P V A V UniRef90_C1P968_353_ L H D V T E T R R L E Q I R S E F V A N V S H E L K T P V T S UniRef90_UPI000210FB4E_174_ M H D I T K E H E L N E M R K E F V S N V S H E L R T P L T S UniRef90_D7AHN4_358_ L K R L E V V R R D F V A N V S H E L R T P V T V UniRef90_D5X969_353_ D I T E I R E L E K M R S E F V A N V S H E L R T P L T S UniRef90_D8PBL4_363_ D I T E L R R L E K I R K D F V A N V S H E L R T P L T S UniRef90_D7UVZ9_356_ D I T Q I K H L E N V R T E F V T N V S H E L K T P V T A UniRef90_C5D651_348_ D I T E L K K L E Q M R K D F V A N V S H E L K T P I T S UniRef90_F4BLP5_336_ D I T E I R R L E K V R T D F V T N A S H E L K T P V T A UniRef90_Q3MGM2_1112_ R D I T E R K Q I E R M K D E F V S V V S H E L R T P L T S UniRef90_B8G4S5_316_ Q N T F I S V I S H E L R T P V S I UniRef90_E0FRK4_227_ N K M A E E L G Q L E E M K N E F I S N I S H E L K T P L T S UniRef90_B1ZT44_154_ D V T R L Q Q L E A I R Q D F V A N V S H E L R T P L S L UniRef90_C9RTX6_347_ D I T E L K R L E Q I R K D F V A N V S H E L K T P V T S UniRef90_A4XMJ6_338_ K L D S M R K Q F V A N V S H E L R T P I T T UniRef90_UPI C5_286_ D I T E L K K L E Q M R K D F V A N V S H E L K T P I T S UniRef90_A5ILS8_180_ R K L D E M R R E F I A T V S H E L R T P L T S UniRef90_B9K7H6_180_ R K L D E M R R E F I A T V S H E L R T P L T S UniRef90_F2I7J7_353_ V V L A Y D I T E I R R L E K V R S D F I A N A S H E L K T P V T A UniRef90_B5YHW7_385_ N R A K S E F L A N M S H E L R T P L N S UniRef90_C0A490_167_ I T R Q K R L E A V R K E F V A N V S H E L R T P L S V UniRef90_D6XVE0_357_ M R D M T E E R L H D K L R K D F I A N V S H E L R T P I S M UniRef90_A6CR52_338_ D I T E L K K L E Q M R K D F V A N V S H E L R T P I T S UniRef90_F5SLG1_353_ E I T A I R R L E K M R T E F V A N V S H E L K T P V T S UniRef90_A4FWN1_406_ L K E L D N L K S E L I A I V S H E L R T P L T S UniRef90_D3URW5_355_ D I T Q I R H L E N V R S E F V T N V S H E L K T P V T A UniRef90_E8R548_187_ F H N I T E L R R L E R M R Q D F V A N V S H E L K T P L A S UniRef90_C9RD18_197_ F V A N V S H E L R T P L T A UniRef90_C1PEW1_349_ V T I I R D M T E E R K L D K L R D D F I A N V S H E L R T P V A M UniRef90_Q1DCL6_267_ V Y T G R A L Y R E A Q L S R M K T D F V S L V S H E L R T P L T S UniRef90_A8F5W9_184_ L N E M R R E F V S N V S H E L R T P L T S UniRef90_Q2RFL3_239_ R D I T D I R R L E Q M R T E F V A N V S H E L R T P L T S UniRef90_D1JAG2_243_ E R L K E L D R L K S D F V S M V S H E L K T P L T A UniRef90_E5WJB7_351_ D I T E L K K L E Q M R K D F V A N V S H E L K T P I T S UniRef90_F6CIV3_222_ L R D I T E R K K L E Q M R T E F V A N V S H E L R T P L T S UniRef90_F6DKA0_218_ R D V T Q R R K L E K M R T E F V A N V S H E L R T P L T S UniRef90_E3Z246_18_ D I T Q I R H L E N V R S E F V T N V S H E L K T P V T A UniRef90_B5YHI6_153_ D I T R L K Q L E N V R R D F V A N V S H E L K T P V T A UniRef90_A9GF59_268_ L R R L E T I R T D F V A N V S H E L R T P V T A UniRef90_E8WRA5_348_ D I S A M K K A D Q I R R D F V A N V S H E L R T P V T V UniRef90_Q097K6_265_ V Y T G R V L Y R E A K L S R L K T D F V S L V S H E L R T P L T S UniRef90_E1KHP7_365_ Q K L D N M R R E F V A N V S H E L R T P L T S UniRef90_A9AVU3_644_ D R M K T E F V A T A S H E L R T P L T S UniRef90_A6UWZ1_410_ L K K L D K F K S E I I S I V S H E L R T P L T S UniRef90_D4Y401_348_ D I T E L K K L E Q M R K D F V A N V S H E L K T P I T S UniRef90_D7ATL0_341_ K L D T M R K E F V A N V S H E L R T P I A T UniRef90_Q1PXZ9_292_ K A V S A L K R A N R M K S E F L A N M S H E L R T P L N A UniRef90_E7M6U2_368_ V R R L E R I R S E F V A N V S H E L K T P V A A UniRef90_D7BCF8_586_ I R D V T P Q K D A E R I K S E F I A A V S H E L R T P L A A UniRef90_D0MGH9_109_ T L E K E I E E L K R M E N Y R R E F L G N V S H E L K T P I F S UniRef90_UPI000212A698_523_ E K L I Q A K E G A E L A S K T K S E F L A N M S H E L K T P L T A UniRef90_E1K386_410_ Q A E E L E K S Y N E L K E L D K L K S D I V A I V S H E L R T P L T S UniRef90_B9M7U2_251_ K E I E Q M K D E M I S A V S H E M R T P L T A UniRef90_Q3AAG9_206_ F D D I T E E R K L E K M R S E F I A N V S H E L R T P L T S UniRef90_UPI0001E8932B_350_ D I T E L K K L E Q M R K D F V A N V S H E L K T P I T S UniRef90_C4G960_243_ L G R M Q S V E A S R Q E F V S N V S H E L K T P L T S UniRef90_C6J6K9_342_ Q D V T E I R K L E R M R S E F V A N V S H E L K T P I A A UniRef90_A0YQV3_1128_ Q L E R A T R L K D E F M A N M S H E L R T P L Y S Fig. S5 The complete multiple sequence alignment (MSA) of HK homologs retrieved from the UniProt database and aligned using the Consurf Protein database. 18 Highly-conserved active site residues are outlined in red and indicated by a red arrow (Asn found in N Box region, Asp found in G1 box region, Leu found in G2 box). S11

12 UniRef90_E3PT03_185_ I E D I T E R I K L E T I R S D F V A N V T H E L K T P L T S UniRef90_B7GGU1_348_ D I T E L K K L E Q M R K D F V A N V S H E L K T P I T S UniRef90_Q39S62_358_ L T R L E N I R R D F V A N V S H E L R T P V T V UniRef90_B8D0H3_363_ L R R L E Q L R K E F V A N V S H E L K T P L T S UniRef90_F5L948_225_ D I T Q L K K L E R I R Q D F V A N V S H E L K T P I T S UniRef90_E0NJE3_365_ R L D V M R R E F V A N V S H E L K T P I T T UniRef90_A7II27_682_ Q L R T L D E L K D D F V S S V T H E L R T P L T A UniRef90_B9DNB3_332_ D I T S L K K L E N L R R E F V A N V S H E L K T P I T S UniRef90_A7HN49_181_ K Q L D K L R R E F I S N V S H E L R T P L T S UniRef90_E6QIY0_358_ R R L E R M K D E F V S T V S H E L R T P L T S UniRef90_F4L5H3_895_ E R L Q Q V D K L K D Q F L A N T S H E L R T P L Q G UniRef90_A9AUY3_448_ I E D V T R E R E I D K M K N E F V S V V S H E L R T P L T S UniRef90_Q1IKU6_785_ Q D V T K R R E V D R M K N E F I S V V S H E L R T P L T S UniRef90_B5YD58_456_ R D I S L A K E L D R M K S E F V A N V S H E L K T P L T A UniRef90_UPI00016BFB3D_364_ M Q D V T S Q Q K L D Q M R K D F V A N V S H E L R T P L T T UniRef90_E8SIU9_335_ D I T Q L K K L E N L R S E F V A N V S H E L K T P I T S UniRef90_B9E797_336_ D I T Q L K I L E Q M R K D F V A N V S H E L K T P I T S UniRef90_Q8EPE4_222_ D I T E L K K L E K M R K D F V A N V S H E L R T P I T S UniRef90_D5DL55_355_ R D M S E E R K L D K S R K D F I A N V S H E L R T P I S M UniRef90_E0HX50_173_ L R D I T K E K E V E A M R R D F V A N V S H E L R T P L T S UniRef90_B8DZU7_455_ R D I S L A K E L D R M K S E F V A N V S H E L K T P L T A UniRef90_D6GSX7_369_ E R L D H M R K E F V A N V S H E L K T P I T T UniRef90_D7CLL1_123_ R K F D Q M R S E F V A N V S H E L R T P L T A UniRef90_Q6LXP6_406_ L K E L D K L K S E L I A V V S H E L R T P L T S UniRef90_Q67LR3_257_ R D I T K E T E L D R M K T E F I A T V S H E L R T P M T S UniRef90_A6LP23_177_ L H D V T Q E R I L E N A R K E F I S N V S H E L R T P L T S UniRef90_B1HWE4_263_ M H D I T E L V R L E Q I R K D F V A N V S H E L R T P I T S UniRef90_Q2B2H9_351_ D I T E L K K L E Q M R K D F V A N V S H E L K T P I T S UniRef90_B8FA30_356_ D M T Q V T K L E N M R R D F V A N V S H E I K T P I T A UniRef90_B5YJP0_360_ R D I T Q E K E V D R L K T E F I T V V S H E L R T P L T S UniRef90_F0JJT0_240_ R L R R E E R M R A D F V A M L S H E I R T P L T S UniRef90_E1KHE7_585_832 - I T E H K E I E S M L I E T N N K L K E L D Q A K T D F F S N V S H E L R T P L T I UniRef90_C9LCS1_237_ N K M L K K M K V L D D S R Q E F V A N V S H E L K T P M T S UniRef90_B7IEX9_176_ L H D V T Q E R V L E N V R K E F I S N V S H E L R T P L T S UniRef90_Q8TMU8_474_720 - I T E R K N Y E I E L F R A K Q D A E V A N R A K S A F L A N M S H E L R T P L N S UniRef90_Q71Y67_354_ R D M T E E K Q L E K M K S D F V N N V S H E L R T P I S M UniRef90_D5DMV7_352_ D I T E L K K L E Q V R R D F V A N V S H E L K T P I T S UniRef90_A5D189_222_ L R D I T E R K I L Q E M R S E F V A N V S H E L R T P L T S UniRef90_C0Z7X8_357_ I R R L E K M R S E F V A N V S H E L R T P I T S UniRef90_D4TMU3_804_ R D I T E R K Q V E K M K D E F V S V V S H E L R T P L T S UniRef90_Q0W6Q5_299_ E R L K S L D R M K M E F F T L I S H E L R T P L T T UniRef90_B9E6M6_350_ K K I D Q M K Q M F I A N V S H E L R T P I Q M UniRef90_UPI000212C3B1_361_ V R R L E R M R S E F V A N V S H E L K T P I A A UniRef90_A6UPU7_401_ E E L K E L D N L K S D L I A I V S H E L R T P L T S UniRef90_A5UQE0_447_ S D V T R E R E A D R L K S E F L S I I S H E L R T P L T S UniRef90_A1HSJ7_400_654 D I T E H K Q L E V K L T E A L A A A E A A N Q A K D Q F L A A M S H E L R T P L N A UniRef90_F2JII5_342_ Q D V T K Q Q K L D Q M R K E F V A N V S H E L R T P L T T UniRef90_Q8R6U6_339_ M H D I T E Q H K L D Q M R K E F V A N V S H E L R T P L A T UniRef90_Q8PT37_279_ N R S M S E F L A T M S H E L R T P L T A UniRef90_D8G5S6_939_ K Q I E R M K D E F I S I V S H E L R T P L T S UniRef90_A8FJB5_372_ E K I D A E R R E F V A N V S H E L R T P L T T UniRef90_A5ITL6_318_ M H D I T N L K Q L E N L R R E F V A N V S H E L K T P I T S UniRef90_F6B3A9_217_ L R D I T E R R K L E R M R T E F V A N V S H E L R T P L T S UniRef90_D9QVY1_356_ R D V T E L R R L E Q I R T E F V S N V S H E L R T P L T S UniRef90_A9AA11_406_ L K E L D N L K S E L I A I V S H E L R T P L T S UniRef90_D5E8Z5_365_ E Q A N R T K S E F L A N M S H E L R T P L N S UniRef90_D2QCT5_672_ E R L K Q S D E Q K D D F V S T V T H E I R T P L T S UniRef90_A3DHV5_362_ L H D I T E Q Q K L E N M R K E F V A N V S H E L R T P L T S UniRef90_B0ADS6_194_ I E D I T E L V K L E N M R K D F V A N V S H E L K T P L T S UniRef90_E7RC70_372_ E K I D I E R R E F V A N V S H E L R T P L T T UniRef90_A0B593_141_ L K E L D S I K S E F V S M V S H E L R T P L T V UniRef90_D4IVS0_239_ I K K L Q A I D Q S R Q E F V S N V S H E L K T P L T S UniRef90_B1YMQ3_615_ D R M K E E F V S T V S H E L R T P L S S UniRef90_C1F3B6_554_ Q D I T E R Q R L D R M K D E F I S T V S H E L R T P L T S UniRef90_E7RJV0_231_ Q D I T E L K K L E Q I R K D F V A N V S H E L R T P V T S UniRef90_D3UPQ2_354_ R D M T E E K Q L E K M K S D F V N N V S H E L R T P I S M UniRef90_E0RD24_357_ Q D I T A I R R L E N M R S E F V A N V S H E L K T P I A A UniRef90_F4LRR4_346_ R D I T D L R K L E K V R T E F V A N I S H E L K T P L T S UniRef90_Q7NDS1_97_ Q E L D Q A Q T D F V S T V S H E L R T P L T S UniRef90_Q647M8_182_ L E E V T R M K T D F L S I T S H E L R T P L T P UniRef90_B8G6G1_479_ D R L K D E F I G I V S H E L R T P L T S UniRef90_B7GKK3_359_ R R L D K L R K D F I A N V S H E L R T P I A M UniRef90_C1DVW6_110_ Q A K K D F V S N V S H E L K T P I S V UniRef90_C4L472_342_ D I T E L K R L E K M R K D F V A N V S H E L K T P L T S UniRef90_D7GPR0_250_ T I N K T L E K L K A V D Q S R Q E F V S N V S H E L K T P I T S UniRef90_B4S8R4_490_ F R D I T E Q K K S E R L I K E N I R L K N D F I A N V S H E L R S P L F S UniRef90_A5GAH6_406_ D I T E R K E M E Q M K D E M I S A V S H E M R T P L T A UniRef90_C6WZH9_504_ V G T S T D I D D M K R Q E Q Q K D D F I K M A S H E L K T P V T T UniRef90_B0TGE0_245_ V R R L E Q M R T E F V A N V S H E L R T P L T S UniRef90_D5E978_387_ V K T T K T E F I A T M S H E L R T P L N A UniRef90_F5LRU7_361_ M R R L E R M R S E F V A N V S H E L K T P I A A UniRef90_Q4A159_364_ L H D V T E Q Q Q V E R E R R E F V A N V S H E L R T P L T S S12

13 H Box Input_pdb_SEQRES_2C2A I K A Y A E T I Y N S L G E L D L S T L K E F L E V I I D Q S N H L E N UniRef90_A5IIT0_236_493 I K A Y A E T I Y N T L E E L D L G T L K E F L E V I M D Q S S H L E N UniRef90_B9KAB5_220_473 I K A Y T E T M Y N S L E E L D T D T L K E F L E V V L D Q S N H L E N UniRef90_UPI000210F96E_244_482 I R A Y V E T I L N S L D E L N K T M L K D F M K T V F D E T L H L E N UniRef90_B7IDY9_224_462 I K A Y T E T L L N M E V D R E S Q R E F L E T V Y E Q S E R L E S UniRef90_A6LN40_223_461 V K A Y T E T L L N M E I D P E S Q K E F L S I I Y E Q S E R L E S UniRef90_A8F6Z9_244_486 I K A Y T E T I L A D P E S M D K D T L T G F L Q I V Y K E S L H L E S UniRef90_A7HK36_224_466 I K A Y V E T M L N M P M S Q E E I H E F L D V V Y S Q S I R L E E UniRef90_D5WTD2_380_614 I C G L A E T V V E E D L A - - P E E R R H F L E L I H R E A R R L E Q UniRef90_Q67RF9_205_438 I Q G F A E T L L E G A L D - E P E T A R H F V E I M L R E S R H L G A UniRef90_B3E3A4_350_579 I K G Y G E T L L D G A L E E S P E R S R R F V E I I V S H A E R L T N UniRef90_C1P968_353_593 V K G F A E T L L D G A M Y - D E A T L R E F L K I I Y D E S D R L H R UniRef90_UPI000210FB4E_174_410 I H G Y A E T L L N D P D I - D P E T R Q R F L S I I E N E A A R M T R UniRef90_D7AHN4_358_582 I K G Y A E A L A G G L V E E D P E R A G R F L E I I C S H S E R L A D UniRef90_D5X969_353_585 I K G F V E T L L D G A M E - D R E V A R R F L E I I N V E T N R L S R UniRef90_D8PBL4_363_603 I K G Y V E A L L D G G K D - E P A T A T A F L E I I M R Q S N R L N L UniRef90_D7UVZ9_356_591 L K G F A E T L L D G A M Y - D E A L L K K F L G I M K D E S D R L H R UniRef90_C5D651_348_580 I K G F A E T L L D G A M K - D E Q T L E Y F L S I I W K E S E R L Q T UniRef90_F4BLP5_336_567 L K G F S E T L L D G A M E - D K E V L K Q F L E I M L A E S S R L D F UniRef90_Q3MGM2_1112_1344 I H G S L G M L A S G L L P A D S E Q G R R L L E I A T D S T E R L V R UniRef90_B8G4S5_316_540 I K G F A E T M L R P D G Q F T V E Q Y R E A L Q V I G E E A D R L A R UniRef90_E0FRK4_227_461 I K G F A I T A M D L V E K N S E L Y E Y L N I I D E E T D R L S R UniRef90_B1ZT44_154_390 I K S A A E T L L D G G K N - D P A V N A R F L E I I D K H A N R L S L UniRef90_C9RTX6_347_579 I K G F A E T L L D G A M K - D E A A L E H F L T I I L K E S E R L Q T UniRef90_A4XMJ6_338_561 I K T Y S E T L L D V D - N E E T K K Q F L S V I I K E C D R M T R UniRef90_UPI C5_286_519 I K G F S E T L L D G A L E - D R D T L E Y F L N I M L T E S D R L Q S UniRef90_A5ILS8_180_410 I H G Y A E T L L E D D L E - D K E L V K R F L K I I E E E S A R M T R UniRef90_B9K7H6_180_412 I H G Y A E T L L E D D L E - N K E L V K R F L K I I E E E S A R M T H UniRef90_F2I7J7_353_596 L K G F T E T L L D G A L E - D E D T A R E F V E I M N R E A N R L G F UniRef90_B5YHW7_385_611 I I G F T E V L Q D Q L F G T L N E K Q L E Y L K D I H D S A K H L L N UniRef90_C0A490_167_399 I K G Y A E T L I D G G P D M P P A Q R D R F L H I I R R H T D R L N T UniRef90_D6XVE0_357_597 L Q G Y S E A I I D D V A G - S D E E K K E L A Q I I Y D E S L R M G R UniRef90_A6CR52_338_570 I K G F S E T L L D G A M N - D K E T L E M F L N I I L R E S D R L Q S UniRef90_F5SLG1_353_585 L R G F A E T L L D G A A E - D P D M R K E F L E I I Q A E S L R L E R UniRef90_A4FWN1_406_639 I K G Y V E L V L D G T M G A I N D S Q R K C L Q V A D D N I V R L R R UniRef90_D3URW5_355_589 L K G F A E T L L D G A M Y - D E A L L K K F L T I I K E E S D R L H R UniRef90_E8R548_187_428 I R A Y A D S L L D W A L E - D P E I T R R F V S Q I D E Q A E R L D V UniRef90_C9RD18_197_417 I K G F V E A L E D G A L E - D R E T A Q E F L Q I I A S E T E R L I H UniRef90_C1PEW1_349_587 L Q G Y S E A I I D D I A A - S E E E K K E F A K I I Y D E T L R I G R UniRef90_Q1DCL6_267_509 I R M F I E T L A L G R L K - D P A Q T Q E V L T L L M R E T E R L S I UniRef90_A8F5W9_184_414 I H G Y A E T L L N D P D M - D A E T R D R F L K I I E N E S A R M S R UniRef90_Q2RFL3_239_485 I R G F V E T L L E G A L E - D P E V S R R F L G I I N H E A Q R L Q Q UniRef90_D1JAG2_243_467 M R T S A Q V L E A A G I A T E T K R E M L D I I L R N I D R Q T N UniRef90_E5WJB7_351_584 I K G F S E T L L D G A M E - D K Q A L N D F L S I I L K E S D R L Q S UniRef90_F6CIV3_222_456 I K G F L E T L L D G A M N - D P K T S R Q F L E I M S Q E T E R L T R UniRef90_F6DKA0_218_449 I N G F M E T L L D G A I D - D P V I A R R F L E I M N T E S N R L S R UniRef90_E3Z246_18_252 L K G F A E T L L D G A M Y - D E M L L K K F L T I I K E E S D R L H R UniRef90_B5YHI6_153_383 I K G Y A E T L L D G A I N - D K E N S K K F I E I I K N Q A D R L N A UniRef90_A9GF59_268_496 I S T A A E T L Q L G A L K - D P H E A A E F V D V I D R H A K R L R H UniRef90_E8WRA5_348_584 I K G Y A E T L L S G D L A D D P G R R D R F L G I I Q N H A D R L S D UniRef90_Q097K6_265_506 I R M F I E T L A L G R L K - D P A Q T Q E V L Q L L S R E T E R L S A UniRef90_E1KHP7_365_592 I K S Y S E T L M D G A L E - D K E T A Y R F L N V I N S E A D R M T R UniRef90_A9AVU3_644_869 I S G Y I D L L M L N T L G P L T E Q Q R Q F L S V V K N N I E R L N A UniRef90_A6UWZ1_410_640 I K G Y V E L V L D G T M G T I N E S Q K R C L E I A N E N I D R L K R UniRef90_D4Y401_348_580 I K G F A E T L L D G A M K - D E R T L E H F L S I I W K E S E R L Q T UniRef90_D7ATL0_341_566 I K S Y V E T L L Y S D V - - D A E Y S K K F L K I I D S E T D R M T R UniRef90_Q1PXZ9_292_526 I I G F A E V L K D K L C G E L N A E Q E D F V K D I H S S G R H L L Q UniRef90_E7M6U2_368_603 V K G F A E T L M A G A L E - D K E M A R S F L Q I I Y D E S D R L N R UniRef90_D7BCF8_586_818 I M G F A E L L T S G E I P L E E G Q E F L R I I Y D N G R R L K N UniRef90_D0MGH9_109_352 I R G F T E T L L E A D P G - D E A T R R A F L E K I L R N A D R L A N UniRef90_UPI000212A698_523_767 I I G F S E L L N T R M F G E L N E K Q L G F V E Y I I K N G N H L L E UniRef90_E1K386_410_648 I K G Y V E L V L D G T M G A I T E S Q K K C L E I A N K N I D R L K R UniRef90_B9M7U2_251_475 M L G F T E Y L L E N E V D P S Q L K N Y L N T I Y K E T A R L K E UniRef90_Q3AAG9_206_440 I K G F L E T L L D G A L E - D K T I A K H F L Q I M N S E T E R L T R UniRef90_UPI0001E8932B_350_584 I K G F S E T L L D G A M N - D K D T L E Y F L S I I L K E S D R L Q S UniRef90_C4G960_243_473 M K V L A D S I N S M E G A - P L E L Y Q E F M Q D M S H E I D R E T K UniRef90_C6J6K9_342_579 V K G F A E T L L A G G V K - D E E T T R S F L Q I I Y D E G D R L N R UniRef90_A0YQV3_1128_1360 I L G L S E V L Q Q E V Y G V L N A E Q L R S L S T I E Q S G Q H L L E S13

14 UniRef90_E3PT03_185_421 I N G F V E T L K S N A G I - N K A M R T K F L D I I E V E S N R L Q R UniRef90_B7GGU1_348_587 I K G F A E T L L D G A M H - D A Q T L E Y F L T I I L K E S E R L Q H UniRef90_Q39S62_358_588 I K G Y A E A L I D G A M E T D P E R A R K F V G I I L S H S E R L A A UniRef90_B8D0H3_363_592 I I G Y I D T I I D N D I K - D D T T I K R F L S I I K D E A D R L Y L UniRef90_F5L948_225_461 I K G F A E T L L D G A M Y - K E E H L K H F L S I I H K E A E R L H R UniRef90_E0NJE3_365_590 I G S Y T E T M L D V D M - - D L D S I R N F L R V I D R E N N R M A R UniRef90_A7II27_682_911 I R A L S E L M L D A P D M E E A Q R Q E F L A I I V G E S E R L G R UniRef90_B9DNB3_332_557 I K G F A E T L I D G A K N - D E N S L D E F L N I I L K E S N R I E S UniRef90_A7HN49_181_412 I H G Y A E A L L D D D L S - N K E L V R K F L G V I E S E S A R M T R UniRef90_E6QIY0_358_593 L R A S L G L I A S G A L E K R P E K Q K Q M I E V A L A N S D R L I R UniRef90_F4L5H3_895_1123 I I G L S E A L L E D E L D P E R K A N L T M V V S S G K R L N S UniRef90_A9AUY3_448_692 I L G Y T E L L L A R E F K P V E R Q E F V Q T V Y D Q A N Q L S K UniRef90_Q1IKU6_785_1018 I R G S L G L L A G G A L R K D P E K A D R M L D I A L K N T E R L V R UniRef90_B5YD58_456_687 I K G Y S E L L I K M N L P P E K V R N Y Y Q I I Y K E S E R L T Q UniRef90_UPI00016BFB3D_364_592 I K T Y T E T L I S G A I D - E K E T A L D F L S V M E K E T D R M T T UniRef90_E8SIU9_335_566 M K G F T E T L I D G A K N - D E A S L D L F L N I I L K E S N R I Q S UniRef90_B9E797_336_561 I K G F T E T L L D G A K E - D K D T L E M F L D I I L K E S N R I Q V UniRef90_Q8EPE4_222_453 I R G F A E T L L D N N I T - D P A T - K E F M E I I Y K E S H R L Q L UniRef90_D5DL55_355_590 L Q G Y S E A I I D D I A S - T D E E K K E I A Q V I Y D E S L R M G R UniRef90_E0HX50_173_410 I H G Y A E T L A E D D L E - D K E T V Y R F L S I I E N E S A R M T R UniRef90_B8DZU7_455_686 I K G Y S E L L M K M N L P P E R V R N Y Y Q I I Y K E S E R L T Q UniRef90_D6GSX7_369_596 I R T Y S E T L L D G A L K - D P A I A K K F M E V I V K E S D R M T S UniRef90_D7CLL1_123_351 I K G F V E T L L D G A L E - D K L I C R R F L T I I E G E N N R L T R UniRef90_Q6LXP6_406_633 I K G Y V E L V L D G T M G A I N D S Q K K C L Q V A D D N I V R L R R UniRef90_Q67LR3_257_496 I K G A L G L V L G G A A G D L P P E A R E L L T I A R N N T D R L I R UniRef90_A6LP23_177_414 I H G Y A E A L L E D N L D - D K E L I R R F L T I I E S E A A R M T R UniRef90_B1HWE4_263_498 I K G F S E T L L D G A Y K - D E K M L I S F L E I I Y K E S N R L Q M UniRef90_Q2B2H9_351_585 I K G F S E T L L D G A M H - D Q A A L S A F L D I I L K E S D R L Q S UniRef90_B8FA30_356_583 I K G F V E T L L D G A L D - D K E N A K R F L E I I S R H T D R L K A UniRef90_B5YJP0_360_619 I L G F I E I I N K K F T E N I L P H L D M N N F R L S K A V N K I N K N F K I I L S E G E R I T S UniRef90_F0JJT0_240_468 V R E A V D L V G S G T F G E V N E K Q K R F L D I A G Q E S E R L S D UniRef90_E1KHE7_585_832 I L G Q V E A V L S G Q Y Y K N T L E P N S D I F K T I Q S N A L R L L K UniRef90_C9LCS1_237_475 I K V L A D S L A G Q E D V - P V E L Y K E F M Q D I A V E I D R E N K UniRef90_B7IEX9_176_421 I H G Y A E A L L E D D L E - D K E L I R K F L S I I E S E A A R M T R UniRef90_Q8TMU8_474_720 I I G F S D L L Y E K V Y G E L N L K Q T K A V G N I S N S G K H L L N UniRef90_Q71Y67_354_593 L Q G Y S E A I I D G V A Q - S D E E V R E F A Q I I Y D E S L R I G R UniRef90_D5DMV7_352_585 I K G F S E T L L D G A M E - D P Q L R Q Q F L S I I L T E S Q R M E S UniRef90_A5D189_222_456 I R G F A E T L L D G A L E - E P D T A R R F L E I I N S E T E R L S R UniRef90_C0Z7X8_357_585 I K G F T E T L L E G A M Q - D E E T C R N F L Q I I S D E S E R L Y R UniRef90_D4TMU3_804_1043 I H G S L G M L T S G L L S T T S E Q G K R L L Q I A T D S T E R L V R UniRef90_Q0W6Q5_299_531 I K G Y A E L L K D G T L G P V N D E Q R D R L S R I D A S V D R L T G UniRef90_B9E6M6_350_585 L Q G Y T E A I L D G I V S - E K S D V D E F L N I I L D E S K R L N R UniRef90_UPI000212C3B1_361_596 V K G F A E T L L A G A L N - D K E T A R S F L Q I I F D E S E R L N R UniRef90_A6UPU7_401_631 I K G Y V E L V L D G T M G T I N E S Q R K C L Q V A D D N I I R L R R UniRef90_A5UQE0_447_687 I M G Y T E L L L A R E F S P A E R R E F V Q T V Y N E A N H L Y Q UniRef90_A1HSJ7_400_654 I M G F S E V L L D Q H F G P L N D K Q Q V Y V N D I L D S A R H L L E UniRef90_F2JII5_342_577 V K S Y T E T L L D G A I D - E K E T A M H F L G V M E K E A D R M T A UniRef90_Q8R6U6_339_575 I K S Y V E T L L Y N D V - - D T E Y S K K F L R I I E T E T D R M T R UniRef90_Q8PT37_279_503 I I G F S E L M L G G S T G E F D E L N K K F L G N I S T S G K H L L S UniRef90_D8G5S6_939_1165 I H G S L G M L A S G L L K A D S E E G K R M L Y I A V D S T D R L V R UniRef90_A8FJB5_372_608 M R S Y L E A L A E G A I G - D K E L A P R F L S V T Q N E T E R M I R UniRef90_A5ITL6_318_551 I K G F A E T L I D G A K N - D A E S L D M F L N I I L K E S N R I E S UniRef90_F6B3A9_217_450 I N G F L E T L L D G A I E - D P K T A R H F L E I M N A E T K R L A N UniRef90_D9QVY1_356_592 I K G Y V E T L L D E R D C - E P G V R E R F L Q V I K D E T D R L E R UniRef90_A9AA11_406_633 I K G Y V E L V L D G T M G A I N D S Q K K C L Q V A D D N I V R L R R UniRef90_D5E8Z5_365_588 I I G Y S Q I L N Q N P S G N L D E K E L K Y S H N I L N S G E H L L E UniRef90_D2QCT5_672_903 I R A L S E I L H D Q A D M D E S M R Q E F L S R V I R E T E R L S R UniRef90_A3DHV5_362_596 I K S Y A E T L L D G A L E - D R E L A G K F L S V I N S E A D R M T R UniRef90_B0ADS6_194_431 I T G F V E T L K I N D D I - D K N T R N H F L D I I E K E S N R L K G UniRef90_E7RC70_372_606 M R S Y L E A L A D G A W Q - D P D L A P N F L N V T Q T E T E R M I R UniRef90_A0B593_141_369 I N S Y I E M F E D G M L G D L T D V Q K E K L Q L I R S Q T D T M I Q UniRef90_D4IVS0_239_469 M K V L A D S L N G S E D V - P I E M Y K E F M V D I G D E I D R E T K UniRef90_B1YMQ3_615_835 I Y G F T E L M L N R E I D P P K Q R K Y L T T I H S E T G R L T T UniRef90_C1F3B6_554_791 L R A A L G L I A G G A L Q R R P E K V Q Q M F D V A I G N C D R L V R UniRef90_E7RJV0_231_462 I K G F S E T L L D G A Y K - D T E T L L S F L E I M H T E S N R L E M UniRef90_D3UPQ2_354_596 L Q G Y S E A I I D G V A Q - S D E E V R E F A Q I I Y D E S L R I G R UniRef90_E0RD24_357_594 V K G F A E T L L G G G V K - D E E T A R S F L Q I I Y D E S E R L N R UniRef90_F4LRR4_346_580 I A G F V E T L L D G A Y K - S Q D H C K Y F L G I I K Q E T D R M T R UniRef90_Q7NDS1_97_331 I K G F V D T L L R S G S Q L S E A Q H R R F L R I I K N Q A D R L T R UniRef90_Q647M8_182_410 M K S Q L Q M L Q E G Y M G K L D K K Q E K S I E V V L R N L T R L E N UniRef90_B8G6G1_479_712 I L G Y T E I L L N R Q D L D S E S Q R E F L L T V S N E A D R L H K UniRef90_B7GKK3_359_585 L Q G Y S E A I V D D I A A - T D E E K K E L A K V I Y D E S L R M G R UniRef90_C1DVW6_110_330 L K S A V E T I E E E K D P V M I K K F V N I A K K R I E Q M D H UniRef90_C4L472_342_574 I R G F S E T L L D G A K E - V P E L R D Q F L D I I Q K E A T R M Q M UniRef90_D7GPR0_250_485 I R V L A D S L M G M E D T - P K E L Y Q E F M H D I S D E I D R E S K UniRef90_B4S8R4_490_736 I L G F S S T L L K E R K E L D F E T T G E F L G I I H D E S K R L S S UniRef90_A5GAH6_406_635 M L G Y T E F M L D N E V P A D Q Q R E Y L R I T L H E S E K L N E UniRef90_C6WZH9_504_741 I K G Y V Q L L K R T R K D S D D K F L V N S L N T I E N Q V N N L S V UniRef90_B0TGE0_245_473 I K G F V E T L L D G A L D - D T K V A R R F L S I I N E E T Q R L Q R UniRef90_D5E978_387_612 I I G F S Q L L S S N R Y G N M N E K E L K Y S F S I L K G G K H L L N UniRef90_F5LRU7_361_595 V K G F S E T L L A G A L D - D K E T A R S F L Q I I F D E S E R L N R UniRef90_Q4A159_364_598 M N S Y I E A L E S G A W K - D G E L A P Q F L S V T R E E T E R M I R S14

15 Input_pdb_SEQRES_2C2A L L N E L L D F S R L E - R K S L Q I N R E K V D L C D L V E S A V N A I K E F A S S H N V N V L F UniRef90_A5IIT0_236_493 L L N E L L D F S R L E - R K S L Q I N R E K I D L C G L I E S A V N A I K E F A S S H N V N V L F UniRef90_B9KAB5_220_473 L L N E L L D F S R L E - R K A L Q I K K E K T N I C E L L E S A V G A I K E F A A S Q G V K V F L UniRef90_UPI000210F96E_244_482 L V N Q I L D F A K I E - Q K T L K L E K T W F D L V E L C Q E V I Q S M G E F A K S K Q V S L E Y UniRef90_B7IDY9_224_462 L L N D L L D F T L I E - S G T M E L E Y S E F D L C E L I K E V V E N L K Q L A E K Y K V K I N V UniRef90_A6LN40_223_461 L L N D L L D F T L I D - S G S M E L E Y S K F N I C N V V D D V L K K L L S F A Q K Q D V K L E K UniRef90_A8F6Z9_244_486 L L D E L L D F A K L E - Q K A M V L E K S K F D L T E L I M Q V I Q S M S E F A R S K K V K L Q C UniRef90_A7HK36_224_466 L L S D L L D F S Q L E - S H T M K I L K E S V N I C E I I E H S I E T L E N M A K E K D V T V E Y UniRef90_D5WTD2_380_614 L V R D L L D L S R L E - S H L T T L H R R P V D L A E M A R D V V Q D Y G Y A A E Q A G T D L Y L UniRef90_Q67RF9_205_438 L I D E L L D L S R V E - S G K F R M Q R R P T V P A E L I A A T A A R F A Q K A E R A G V Q L V T UniRef90_B3E3A4_350_579 L I N D I L T L S K L E - A R D A A L T L H P L D L C G T I R K A Q M L M E D H A R S K G I R I A A UniRef90_C1P968_353_593 L I S D I L D L S R I E - Q H R I L L K M E Q L N V V D V V A E T V Q T M R K R I E K K Q L E L V L UniRef90_UPI000210FB4E_174_410 L I N D L L D L E R L E - S G E A R F E F Q P V D L C S V I K Y V L S I V E P L A V Q Y G V K V E Y UniRef90_D7AHN4_358_582 L I R D L L T L S Q L E - S G G L Q L E L T Q I H L D R A V S H A A G L L E Q K A A R K E I A I D I UniRef90_D5X969_353_585 L I N D I L S L S S I E - A K N K E I S R S P V N F N E I V E K V L P I L V P M A E E K N I T V E T UniRef90_D8PBL4_363_603 I L D D L L Q L S Q I E - S G Q V L F R R E P V D L R A L L E R T V A V I K P L A D K K H H T I E L UniRef90_D7UVZ9_356_591 L I L D I L A L S R I E - Q H A L P I N Q E V V D I Y Q T I T E A A Q T V T E S A K Q K N I D L V L UniRef90_C5D651_348_580 L V Q D L L D L S K I E - Q Q G F R L H L E T V D L A D L L R E I A V M F Q Q K S E E K G I D F Y L UniRef90_F4BLP5_336_567 L V N D I L E L S K L E - Q K Q V P M N M Q E V N L T E A V L S T F Q L V K Q T A D E K E M K L N L UniRef90_Q3MGM2_1112_1344 L I N D I L D I E R I E - S G R V K M E R E S C N L T D L I E S A V S I M R P L A N K A G V K L S V UniRef90_B8G4S5_316_540 Q I Q D L L D V S R I A - A G G L R L E Y S D V S L Q L L V K E V V R R F A A Q V G D R - I E F E I UniRef90_E0FRK4_227_461 L V D E L L D F S K I E - L N K I K L S F E E V E L D K L I E D T V A I L R P H A S N D G V N L I C UniRef90_B1ZT44_154_390 L I D D L L T L S K L D - A E R V E L D I R S V G L R A A V Q D A L D D A L T L A Q P R A I A L E N UniRef90_C9RTX6_347_579 L V E E L L D L S K I E - Q H G F Q L Q L E D V D I A Q V I A E A A A V F R Q K A A E K Q I D L H I UniRef90_A4XMJ6_338_561 L V S D L L Y L S R L D - S G E N I L N L E E V N L S E L V R F V C E K L K I H A K K K N Q T L S C UniRef90_UPI C5_286_519 L I Q D L L D L S K L E - Q H G F K L D V S T F D L S R L L E E V V L M L K G K A E E K E I Q L Q F UniRef90_A5ILS8_180_410 L I N D L L D L E K M E - E S E V N F E M K D V D L C E V I E Y V Y K I I Q P I A E E N E V D L V V UniRef90_B9K7H6_180_412 L I N D L L D L E K M E - E S S V E F E M K E L D L C E V V D Y V Y R I V Q P I A E E N E V E L E I UniRef90_F2I7J7_353_596 L I N D I L D L A K I E - Q D Q L G H R S E T V Q L N R V I A E V V H S L E I P A S E N R V E V H Y UniRef90_B5YHW7_385_611 L I N D I L D L S K I E - E G K T E L E L S E F R V S D V V N V S L I M F K E K A S K H G I K L D A UniRef90_C0A490_167_399 L L E D L L T L S R L E - S P N T G L R R E P G D L D R L V K T L L D E Y R T R P A A A R H T L Q L UniRef90_D6XVE0_357_597 L V N E L L D L A R I E - A G H I R L N L E P V N M T E F V E R I S R K F Q G V A D D N N V E L M G UniRef90_A6CR52_338_570 L I T D L L D L S K M E - K Q G F Q L N V H E T D M K E L L I E V I T I L E K K P E K K D I E L V L UniRef90_F5SLG1_353_585 L I A D L L D L S K I E - S R N M P L D I Q T I G I G E L L R S T A K T V E E Q M R K R K L S F R V UniRef90_A4FWN1_406_639 L I E S M L D L S K I E - R G E L E M Y R E K V N I C E I V G D V I Q Y L K P L A T E K N I K L N K UniRef90_D3URW5_355_589 L I M D I L A L S R I E - Q N P V T E N V G T V D V D D V I D Q S V R T I F E L A T E K N I Q V S A UniRef90_E8R548_187_428 L I H D L L S L A R I E - S G Q D L I T F R P L A L A P L V E R R V E S F R D R A Q S R N V S L S F UniRef90_C9RD18_197_417 L V E D L L K L S R L E - N R Q T F L R R Q E V N L N E L I H N I A L V W R R R A E E K G L A F E V UniRef90_C1PEW1_349_587 L V N E L L D L A R M E - A G H F S F R M E P V S F N S F I E R V T H K F N G I A K E K G I D L V C UniRef90_Q1DCL6_267_509 F V E R V L D W A R I E - G G R K V Y Q R E I V P V S E L V N A A V E A F R T Q R M E D G V D L T V UniRef90_A8F5W9_184_414 L I N D L L D L E R L E - S G E A R F D F Q K I D L C S V V K Y V V S I T E P L S I D Y D V K V E Y UniRef90_Q2RFL3_239_485 L I E D L L S L S R L E - S Q P K R Q D A G R A D L A A T L D R V L T T V N Q L A R E K G V A L E K UniRef90_D1JAG2_243_467 L V N D L L D L S R I E - S G R M K L K F E S V S L D S L I A D S I E S V K H A A S E K G I K L N V UniRef90_E5WJB7_351_584 L I Q D L L D L S K I E - K Q G F S L S I Q Q L D L T D V L E D V M A I M K G K A A E K E I V F K Y UniRef90_F6CIV3_222_456 L V D D L L D L S K I E - E R R V V H R W Q P V N L V D I I N R V A S L F R P Q A K E K A L T L S L UniRef90_F6DKA0_218_449 L V D D L L Q L S K L E - Y R K G N L N K Q P V N L T E V I R D T V E V F K N R A A E K N L S F T S UniRef90_E3Z246_18_252 L I M D I L A L S R I E - Q N P V A E N I E P V D V D E V I E Q S S R T I F E M A T E K N I Q V S V UniRef90_B5YHI6_153_383 L V E D L L T L S R I E - F G D I K I E K E N I V L D E L I S S V F Q I L K D K A Q K K G V L L Q K UniRef90_A9GF59_268_496 L V D D L L D L S K I E - A K N F R L A L S E L D I A P A I E H V T Q L L A E A A R R R R V T L T V UniRef90_E8WRA5_348_584 L V R D L L A L S E L E - S G E L A M Q P Q K V L I E D A V R Q A L L L L S P R A E E K G I V M E L UniRef90_Q097K6_265_506 L I E R V L D W A R L E - S G R K E Y Q R E P T A V A D V V D T A V A A F R A Q R L D G G V E L K V UniRef90_E1KHP7_365_592 L V K D L L Q L S R L E - N E Q M Q W N M Q P F C F E S L V R S S V E K I E L S A K E K R Q V I E C UniRef90_A9AVU3_644_869 I L N D L L D V S R I E - S G K V R L Q R K P I N L D E L I Q S T V M S I H Q Q W S G K Q I S L A L UniRef90_A6UWZ1_410_640 L I D N M L D L S K I E - R G E L K M D I S K V N L N D I V Q N V V H S L K P L A D G K N I K I I N UniRef90_D4Y401_348_580 L V Q D L L D L S K I E - Q Q E F Q L H M E T V D L T H L L H E I A I M F Q R K A E E K G I D F R M UniRef90_D7ATL0_341_566 L V K D L L L L S K M D - S E D N N L K F E Q K N L N D I V V E A I N R L S I E A Q K K N Q K L I V UniRef90_Q1PXZ9_292_526 M I N D I L D L S K I E - A G K M E L Q Y E S F L V S D A I E D V Y T I L K G L A K K K Q L Q L K T UniRef90_E7M6U2_368_603 L I G D I L E L S K I E - S K R I P L Q F S P V D V E S I V E N S I Q M M K A E A E K K H I T L E S UniRef90_D7BCF8_586_818 M V D N L L D S S R L E - A G R F E V Y K R P V Q L E E T L R G V A D S F T G V A Q L S Q V E F H R UniRef90_D0MGH9_109_352 L V R D L T E I S R L E - T G E L Q L Q P V P F E L P A L V R E V L E S M E P L A A T R Q V S L R C UniRef90_UPI000212A698_523_767 L I N N I L D L S K I E - A G R M Y L D R E I F C L S Y L I E E V L A T M Y P L A A K K K I D I E T UniRef90_E1K386_410_648 L I D N M L D L S K I E - H G E L E M H M E E I N L K E L V E N V V D T L K P L A D E K N I N I I Y UniRef90_B9M7U2_251_475 L I G N F L D L Q Q I N - A R M T T Y R M H P L D V C L L I K D A A G L F A L D Y D S G R L V V Q C UniRef90_Q3AAG9_206_440 L I D D L L S L S K I E - A K K V D F A P K P L M L Q E L I Q K M K L L F K S R L E E K E L S F I I UniRef90_UPI0001E8932B_350_584 L I E D L L D L S K I E - K Q G F R L N P Q Y L E I N E L L E E I F V I L E G K A R E K E I D L V L UniRef90_C4G960_243_473 I I N D L L S L V R M N - K T G I S M N I A S V N M N E L I E S I L H R V R P I A D R Q G V E L V L UniRef90_C6J6K9_342_579 L I G D L L E L S K I E - S K R V P L E Y A P V Q L S E L F D S L Y E V L L P A A E K K S I S M S H UniRef90_A0YQV3_1128_1360 L I N E I L D L S K I E - S G K M E L Q L S A V N L T Q L C Q S S L N F V K P Q A D Q K N I Q L Q F S15

16 UniRef90_E3PT03_185_421 L I D D I L I L S F I E - G N K S H Y S N E P V N F A D T I N E C I N L V S E T I K D K N V T I D F UniRef90_B7GGU1_348_587 L I Q D L L D L S K I E - Q Q G F T L N I S V V D L H E V L K E V I V M L E A K A N D K Q I T L E Y UniRef90_Q39S62_358_588 L I G D L L T L S Q L E - A G S M N L E K T T V R P E Q V A K R A M E L L Q P K A A R K G I T I D C UniRef90_B8D0H3_363_592 L I K D L L D L S K L E - A K R G E V I L Q P G D L N K I V K K I Y L M L Q E Q A E S K D I D L K L UniRef90_F5L948_225_461 L V E D L L Q L S Q I E - Q R R F G L N W Q E V N L T E V L H N S L Q V V R A K A E V K N I E L I F UniRef90_E0NJE3_365_590 L V T D L L Q L S N M D - Y K E T K W N Y E A V D T Y D F L S T N I N S L D M L I K E K H H K V T L UniRef90_A7II27_682_911 L V N Q V L D M A K L E - S G H G E W H N A D V D L R R L V R D A V T A T T E L A R S R G A E I V F UniRef90_B9DNB3_332_557 L V E D L L D L S H I E - Q Q T - E I R T E R V D L S E V T Q T T V N T L Y A Y A Q R K A I E I D A UniRef90_A7HN49_181_412 L I N D L L D L E K L E - S G D A K F N F S D V N M C K V I E R V V T I L E P L A E D Y G V E L Y T UniRef90_E6QIY0_358_593 L V N D I L D F D R V E - K G G M A L N R E V I P V I D L L R R A A D V A H E A A T R A S V T F R F UniRef90_F4L5H3_895_1123 L V N D I L D F S K L K - N A D I E L D L R P I S L H S M A D V V L K N I S P L V A G K S I E L L N UniRef90_A9AUY3_448_692 M V D D L L N L S R L D - A G Q I K L N R W V V S L H Q I I R E I T K Q L N E T L S E K H - R L L I UniRef90_Q1IKU6_785_1018 L I N D I L D I E K I E - S G N I A L N V Q P L D A A D L I S Q A S A T M H A M A D A N K V R L E T UniRef90_B5YD58_456_687 L I N D L L E V S R I E - S G K I E L K K E M V E I R K V I K E R A T F F Q T Q T S K H T I V L Q F UniRef90_UPI00016BFB3D_364_592 L V H D L L E L S K I D - N K Q M Q L N I Q V M D I Y S L L L D T I R A Q A I Q A K C K H H Q I K L UniRef90_E8SIU9_335_566 L V E D L L D L S K I E - Q N T - T L E K H L I D L S D V A K S S F S V I Q P L A N E K S I Q L I D UniRef90_B9E797_336_561 L V S E L L E L S K I E - Q A N - H F N M V K V N L P Q K V F N S V E V V Y P L A E K K N I N F N L UniRef90_Q8EPE4_222_453 L I E D L L A L S R L E - R E D F R L L I D N Y D V R Q M V E E I L P Q L H Q K A E N K N L T F D L UniRef90_D5DL55_355_590 L V N E L L D L A R M E - A G H V Q L H L E S V A M V E F T E R V L R K F Q G L A K E K H I K L S L UniRef90_E0HX50_173_410 L I N D L L D L E K L E - S G E A S F S K E D V E L G E V V N Y V M R I V E P L A S E K N V S I N V UniRef90_B8DZU7_455_686 L V N D L L E V S R I E - S G R I E L K K D M V E I R K I I K E R A T F F Q T R T S K H T I V L Q F UniRef90_D6GSX7_369_596 L I R D L L Q L S H I D - F K K E T W N F D Y T D I N T L I Q D S I D K M E L Y Y Q T K H Q T L V S UniRef90_D7CLL1_123_351 L I D D L L T L S A I E - S R E R K L T L K P V C L V T S I I Q V M N I L G P Q A R E K R L H L E L UniRef90_Q6LXP6_406_633 L I E S M L D L S K I E - R G E L E M Y R E K V N L R S I V C D V I E Y L K P L A T E K N I K L N K UniRef90_Q67LR3_257_496 L I N D I L D I S R I E - A G K M E I R R A P L S P A D V V R R A V E E M N A F A R Q R N I T L T T UniRef90_A6LP23_177_414 L I N D L L D L E K L E - S G N T Q F I F E K I N F T E I L E H V R N I I E P L T K D Y N V D V E F UniRef90_B1HWE4_263_498 L I Q D L L E L S K I E - Q H G F T V N I M P M G L Q D V L I R G A E L T A P R L D E K N M S F Q V UniRef90_Q2B2H9_351_585 L I Q D L L D L S K T E - Q Q G F R L S V Q E M D L S S L L G E C L A I L N G K A K D K A I D I E Y UniRef90_B8FA30_356_583 I V E D L L E L S R I E Q A G G R E L P L E R A S V K E A L E T A V Q V C E A R A I S E N I T V E M UniRef90_B5YJP0_360_619 L I N D V L D I S K L E - S G K A I W N F K E I S I Q G V I T D A Y K A L S S L F E Q K G I P C Y I UniRef90_F0JJT0_240_468 L L T R L L S V S R M E - A E E L H L N P E R V D V A G L V E S T V E R L V P T A R A A S V T L E S UniRef90_E1KHE7_585_832 L I N T L L D F S K I Q - A G K M T L N R Q P V N I N R F L S S Y I S A V K S S A S Q K N I N M I F UniRef90_C9LCS1_237_475 I I T D L L S L V K M D - K K A S E L N I E K V N I N E L M E N I L K R L K P I A D K K K V E L V L UniRef90_B7IEX9_176_421 L I N D L L D L E K L E - S G N T Q F V F E K I N F T E V L E H V R N I I E P L A K D Y Q V E V E F UniRef90_Q8TMU8_474_720 L I N E I L D L S K V E - A G S F E L H Y S T F W L A E V F A E V R D M I F P F A T S K G L K I E L UniRef90_Q71Y67_354_593 L V N D M L D L A R M E - A G F N Q M D N Q K L P L A P L L R K V I S N F D V L A K E N Y V E L G L UniRef90_D5DMV7_352_585 L I Q D L L D L S K V E - Q Q G F K L S I H S V D L N T L L Y E V T T V L S N R A T E K E I T L T L UniRef90_A5D189_222_456 L I D E L L N L S R L E - S H K W V P K R Q P V N M G E L I K R A V A I L Q P R A V E K N L A I K I UniRef90_C0Z7X8_357_585 M I R D I L D L S K I E - Q K R I P L H L G Q V H L Q D L I S S A V A I M H D Q A Q R K E L T I T L UniRef90_D4TMU3_804_1043 L I N D I L D I E R I E - S G K V K M E R E N C D L R D L I N S A I N I M Q A L A D K A G V S L S I UniRef90_Q0W6Q5_299_531 I V E S L S D L S G V A - S R Q Y A G E K I P V S L N E L I E E V V R G I D F L A D L K K L K I T L UniRef90_B9E6M6_350_585 L V N E L L N V A R I D - A G E Q V L N L E A T D I E E L I E R T I M N F K H V A E K H E T E L Q T UniRef90_UPI000212C3B1_361_596 L I G D I L E L S K I E - S K R I P L Q F S P V H L E P F I G N C V H V M N T E A K K K G I E L E L UniRef90_A6UPU7_401_631 L I E S M L D L S K I E - R G E L E M Y R E A M N V K D T V S D V I E Y L T P L A T E K N I K L K Q UniRef90_A5UQE0_447_687 I V E D L L G V T R L E - A G N V R L N Q W A V S L R Q I V S D L T A Q L N N Q I S G K H T M L - I UniRef90_A1HSJ7_400_654 I I N D V L D L A K L N - A G K T N F Y P E T V D I A K L L R Q T L G I I K D R A A A K N I A V E L UniRef90_F2JII5_342_577 L V Q D L L E L S R I D - N K Q I Q L E F V R L N L K P L L E E V L E A Q Y I H I I K H G H Q L E V UniRef90_Q8R6U6_339_575 L V K D L L L L S K M D - S E E N S L K L E P C D F N E I V K N V V N S L S I E A T K K G H N V I L UniRef90_Q8PT37_279_503 L I N S I L D L S K I E - A G K M E L Q S D Y F S L Q D I F S N T K N I L S P L A L K K N I S M D F UniRef90_D8G5S6_939_1165 L I N D I L D I E R I E - S G R V K M E K Q C C N V E D L I T E A V N V V Q A L A N K V E V T L S V UniRef90_A8FJB5_372_608 L V N D L L Q L S K F D - S K D Y Q F N R E W T N F I R F I S L V I D R F E - M T K E Q H V E F I R UniRef90_A5ITL6_318_551 L V T D L L D L S H I E - Q H T - E L D T D Y M N L S D L T R R I I D N M M T Q A N Q K N I S I H T UniRef90_F6B3A9_217_450 L I D D L L Q L S R L E - D R K T V L N K Q P V D L R D V I N G T I Q M F K A R A E E K G I K L F T UniRef90_D9QVY1_356_592 L I T D L L N L S Q L E - S A S D S F D Q E L V N L N Q V I E N V L T T V M P K A D N K G I D L K V UniRef90_A9AA11_406_633 L I E S M L D L S K I E - R G E L E M Y R E K V N L K G I V C D V I E Y L Q P I A T E K N I K L K K UniRef90_D5E8Z5_365_588 L I N S I L D I S K V E - A G K M E Y E P E R V N L P E T I D D I I G L V K P L A M K K S I D I R F UniRef90_D2QCT5_672_903 L I N Q V L D L E R Y D - S G R Y H L S T E R I S M V D L V A E A V E N V A Q L A R D K Q V T L E I UniRef90_A3DHV5_362_596 L V K D L L Q L S R L D - N N Q M K W D M Q K I S F E D L V R N C V E K V K F E S E E K N Q T L E C UniRef90_B0ADS6_194_431 L I E D I L L L S S I E - N G Q - D L S Y E K V K L F D V F K E V C E I T E Y I A S S K N I T I S Y UniRef90_E7RC70_372_606 L V N D L L K L S K M D - S S E T E L S K E M V E F N V F F N R I I D R F E - M S K S H K V N F M R UniRef90_A0B593_141_369 L V N D M L D I L R I G - S R R L R L K K E L S S M E E L I R S V I A S L S R L A A L K E H T I T F UniRef90_D4IVS0_239_469 I I N D L L S L V R M D - K S G A E L N V S A V N I N E L I E H I L K R L K P I A E K A G I E V V F UniRef90_B1YMQ3_615_835 L V N D F L D V Q R M E - S S E Q R Y Q M A T F D L V Q L A A E L I D F H D A S H T T H E L S F E A UniRef90_C1F3B6_554_791 L V N D I L D F E R L G - S G K M Q L Q Y V Q M P A R D L L Q R A V D L Q H S S A Q K A N I T L R V UniRef90_E7RJV0_231_462 L I N D L L D L S K V E - Q S G F R V N A Q P T N M E A V I K R A R E M I Q P K I D E K S I Q L K L UniRef90_D3UPQ2_354_596 L V N D M L D L A R M E - A G F N Q M D N Q K L P L A P L L E K V M A N F D V L A K E N F V E L G L UniRef90_E0RD24_357_594 L I G D I L E L S K I E - S K R S T L D C S P V H V S S F I E S L L E K L N N V A A K K R I T L H M UniRef90_F4LRR4_346_580 L I N E L L Y F S H I E - K N G T E I I K K P V D M I E V I M K A L S I L Q T A I N E K K H K V N L UniRef90_Q7NDS1_97_331 L V E D I L T V S R I Q - S G R L K N L P Q R L D L G E M I D R V F E N L A Q K Y A A Q R M R R E L UniRef90_Q647M8_182_410 L I G D I L D L S R I E - A G R I K L S F D S M N L N D A V K E A I K M Q E A F A K G K N I E I S A UniRef90_B8G6G1_479_712 M V K D L L D V S R L E - A G V V K L N R W A V G L W Q V I E E L T L Q L H T Q L V Q H K L L I D M UniRef90_B7GKK3_359_585 L V N D L L D L A R M E - A G H L M L H H E Q V P L K P Y V E R I I H K F Q A L A K E K G V H L L V UniRef90_C1DVW6_110_330 L I N D L L I L A R L E - S Q E D Q V K K E R Y D L H K Q I E E I F E D L K H L T E E K E I Q L I N UniRef90_C4L472_342_574 L V E D L L E L S R L E - R D D F Q M E L V S V D L N Q L V D E V C L L L S Q K A S K K S I Q L E A UniRef90_D7GPR0_250_485 I I D D L L T L V R M D - K A S S G L S C S Q V Q I N G L I E M V L K R L R P I A R K R N I E L I F UniRef90_B4S8R4_490_736 L I E D V L N V S R I D - S G K V A Y K K K I I D P A P V V I G A C E S L K M M A S E K S V E F S I UniRef90_A5GAH6_406_635 L I G N F L D L Q R L R - S R K E T V T W R P L P L Q P L L E E T V A L F T R V S E R H H I T L D C UniRef90_C6WZH9_504_741 L I G D L L D I S R M E - N G Y L L L N K H K F S L V E L V T E S I E D I K A S E Q S H E I N F E M UniRef90_B0TGE0_245_473 L I E D L L Q L S R I E - S Q L G R V V E G Q S Y L E P E I Q R V R N L L E P I A A D K R I A L Q V UniRef90_D5E978_387_612 L I N D I L D F S K I H - S N K M E L N I E T I N L K E I C D E I K T F I T P L A T K K G I T V E Y UniRef90_F5LRU7_361_595 L V G D I L E L S K I E - S K R I P L Q L S P T H L Q S I I Q K S L H M M E A E A D K K K I A L S M UniRef90_Q4A159_364_598 L V N D L L Q L S K M D - N E S E Q I T K E I V D F N M F I N K I I N R H E M S A - - K D T T F V R S16

17 N Box Input_pdb_SEQRES_2C2A E S N V P C P V E A Y I D P T R I R Q V L L N L L N N G V K Y S K - - K D - A - - P D K Y - UniRef90_A5IIT0_236_493 E S N V P C P V E A Y V D P T R I R Q V L L N L L N N G V K Y S K - - K D - V - - P D K Y - UniRef90_B9KAB5_220_473 E K K V P C F E A E V D P T R M K Q V L L N L L S N G V K Y S K - - K D - E - - P E K Y - UniRef90_UPI000210F96E_244_482 E G P D Q L I V F A D R M R L R Q V L M N L V S N G I K Y S N - - P E - N - - R Y K Y - UniRef90_B7IDY9_224_462 S C E S I S I S A D K R R I K Q A I H N L V D N S I K F S D - - K S - K - - E E R Y - UniRef90_A6LN40_223_461 E C E D V E I S A D K R R I F Q V I Y N L V D N A I K F N D - - R E - K - - P E R F - UniRef90_A8F6Z9_244_486 V K S K P I Y I V A D Q K R I R Q V L I N L I S N G I K Y S K - - E E - A - - S E K Y - UniRef90_A7HK36_224_466 N C S N L E I K C D R K R I E Q V I V N L L S N A I K F S D - - Q A - K - - E K R F - UniRef90_D5WTD2_380_614 D K E G D C R A E V D P D R V R Q V L S N L I D N A L K Y T P - - A G - G - - R UniRef90_Q67RF9_205_438 E A P D G L P L I N G D P D R L V Q V L S N L V E N A I K Y T P - - A G - G - - R UniRef90_B3E3A4_350_579 E C P D G M P K V L A D Q G Q L E Q V L L N L L D N A I K Y T P - - D G - G - - D UniRef90_C1P968_353_593 P Q K R H V M M E A D K D R L R Q I L L N L V T N A I A Y T P - - D K - G - - R UniRef90_UPI000210FB4E_174_410 S C Q D I V L E A D Q D R L I Q M L V N L V D N A V K Y T S L K E T - G E K Q UniRef90_D7AHN4_358_582 S A L A G A P P V L A D P G R L E Q V L I N L M D N A L K Y T P - - P G - G - - T UniRef90_D5X969_353_585 D I H P D L P V I M A N E D L I K Q V L I N L V D N A I K Y T P - - E N - G - - R UniRef90_D8PBL4_363_603 S L P D E Y W V V E G D E E R L V Q V F I N L L E N A V K Y T P - - D Q - G - - R UniRef90_D7UVZ9_356_591 P H A A K P L P I E T D N D K L K Q I I I N L V S N A V A Y T P - - E N - G - - T UniRef90_C5D651_348_580 H A P K S V Y M E G D A N R I K Q I F I N L I T N A L T Y T P - - K G - G - - K UniRef90_F4BLP5_336_567 I E E D T L F I T V D S S R L K Q I L A N L I N N A V V Y T Q - - D S - G - - E UniRef90_Q3MGM2_1112_1344 S H P S I Q L W V D P D R I V Q T L T N L L S N A I K F S T - - A E - K - - T UniRef90_B8G4S5_316_540 R V P D D M P P V Y A D Y E R L R Q V F T N L I E N A V K Y S P - - N G - G - - T UniRef90_E0FRK4_227_461 K K K I D R V V I N G D V N R L K Q V L V N I I D N G I K A C S - - R G - K - - Y UniRef90_B1ZT44_154_390 Q V P V G L S A K V D A A K L R Q V L G N L I E N A I K Y G R - - E R - G - - R UniRef90_C9RTX6_347_579 E S P S G L V I R G D R N R M K Q I L L N L L A N A I T Y T P - - E H - G - - R UniRef90_A4XMJ6_338_561 S I L Q D I V A M V D R D K I E Q V L I N L I S N A V T Y V Q - - E G - G - - Q UniRef90_UPI C5_286_519 E P I P E L T L E G D M N R F K Q V F I N L I N N A I S Y T G - - P G - G - - H UniRef90_A5ILS8_180_410 E C E D V V V R G N K E R L I Q M L L N L V D N A V K Y T S L K E K - G E K K UniRef90_B9K7H6_180_412 D C E N V T V K G N K E R L I Q M L L N L V D N A V K Y T S L K E K - G E K K UniRef90_F2I7J7_353_596 D N D L E A G I R F T T E E M R L K Q I L T N L I N N A I K Y N K A - E G - G - - Q UniRef90_B5YHW7_385_611 E I S K E A D I T V T A D E R K I K Q V L F N L V S N A V K F T P - - D G - G - - S UniRef90_C0A490_167_399 T L G A A P G P L N F D A L K L T Q V F H N L L D N A L K Y T P - - P G - S - - R UniRef90_D6XVE0_357_597 E I D Q S D T E A Y V D P D R I E Q V M T N L I D N A I R Y T S - - V N - G - - S UniRef90_A6CR52_338_570 H A D E A V L A E V D S F R I K Q V F I N L I G N A I T Y T P - - A G - G - - E UniRef90_F5SLG1_353_585 K S E E E F P V Q V D P D R F S Q I L L N L L S N A M T Y T P - - A G - G - - E UniRef90_A4FWN1_406_639 N V E K I T L E A D K D R I T Q V L T N L I E N A I K F S P - - A N - E - - S UniRef90_D3URW5_355_589 P E K T V P P V I I E T N R D Q L Q Q I L I N L L S N A I N Y T P - - V D - G - - K UniRef90_E8R548_187_428 V T E L P P Q F T I L A D E E A L R Q I L D N L I D N A I K Y S S - - D V - D P - W UniRef90_C9RD18_197_417 D L P P G L P L V L G D P E L L T Q V F V N L I D N A L K Y T P - - V - - G - - R UniRef90_C1PEW1_349_587 E T S D S D L T L E I D A D R I E Q V L T N L I D N A L R H T S - - S N - G - - Y UniRef90_Q1DCL6_267_509 E V S D G L P A L D V D R A A V A G A L L N L L Q N A Y K Y S G P - D N - R - - R UniRef90_A8F5W9_184_414 D C E E V T I D G D Q D R L I Q M L L N L A D N A V K Y T S L K E S - G E K K UniRef90_Q2RFL3_239_485 E I P A E I P E L A I S E S Y L N Q V L L N L I D N G I K Y T P - - A G - G - - R UniRef90_D1JAG2_243_467 K L P E G L S S V K G D R E K L T Q V V I N L L N N A I K F T P - - R S - G - - E UniRef90_E5WJB7_351_584 Q R D E K P V Y I E G D I H R L K Q V F I N I I S N A I S Y T P - - N Q - G - - V UniRef90_F6CIV3_222_456 E V P R D L P S V Y G D P D M L A Q V L I N L L D N A I K Y T P - - P R - G - - S UniRef90_F6DKA0_218_449 D I P E L P E V P G D Q G L L V Q V M V N L V D N A I K Y T P - - E G - N - - S UniRef90_E3Z246_18_252 P E K T I P P I I I E T D R D K L Q Q I L I N L L S N A I N Y T P - - V D - G - - K UniRef90_B5YHI6_153_383 E I P P E T A I Y A D Q Y R M S Q I M I N L V D N A I K F T E K - G - - F UniRef90_A9GF59_268_496 D A S A L P P V R C D R R A L E Q V L M N L L D N A I K Y A G - - E G - A - - H UniRef90_E8WRA5_348_584 G S C A P V S A M A D R G R L D Q V L I N L L E N A I K Y S G - - Q G - G - - R UniRef90_Q097K6_265_506 D V P S G L P Q V E V D R H A V A G A L L N L L Q N A Y K Y S G - - Q D - K - - R UniRef90_E1KHP7_365_592 F S I G D K P E V Y A D K D R I E Q V V L N V L T N A I K Y T P - - D K - G - - K UniRef90_A9AVU3_644_869 D V P D D L P P M I A D P E R M R Q I V T N L I S N A Y K Y T R - - D G - G - - R UniRef90_A6UWZ1_410_640 K V E P I T A N L D K D K I T G V L T N L I E N A I K F S P - - V N - E - - N UniRef90_D4Y401_348_580 H A A K S I Y M E G D A N R L K Q I F I N L I T N A L T Y T P - - K G - G - - Q UniRef90_D7ATL0_341_566 D L Q E T P R Y V Y I D R D K M E Q V I V N L V T N A I K Y T P - - E N - G - - M UniRef90_Q1PXZ9_292_526 V I Q E D V K D I E A D R V K F K Q I L Y N L L S N S I K F T P - - Q N - G - - T UniRef90_E7M6U2_368_603 C V E N E L Y I E A D E D R L R Q I L I N L L S N G I S Y T P - - E G - G - - R UniRef90_D7BCF8_586_818 E I P P L P P V E A D P D R I G Q V M G N L L S N A F K F T P - - K G - G - - R UniRef90_D0MGH9_109_352 C V P E N L P H V L G D R E R L R Q V L I N L V D N A I K Y N N - - P G - G - - F UniRef90_UPI000212A698_523_767 V D E V K N D R V F A D R T K L K Q I M F N L L S N A I K F T P - - E A - G - - K UniRef90_E1K386_410_648 K I N D I I M K G D K D R I T Q V L T N L I E N A I K F S P - - V N - G - - K UniRef90_B9M7U2_251_475 P R N I P P L Q G D E A R L H Q V L T N L I S N G L K Y S Q - - A P - A - - R UniRef90_Q3AAG9_206_440 S L P E N L P L V L A D G D M I S Q V L I N L I D N A I K Y T P - - A G - G - - K UniRef90_UPI0001E8932B_350_584 S K P K K E L Y L F A D A S R I K Q V F I N L I N N A I A Y T P - - A G - G - - E UniRef90_C4G960_243_473 E S F R P V F C E I D E T K F S L A I T N L V E N A V K Y N N - - P G - G - - W UniRef90_C6J6K9_342_579 D V P E H L F I E A D E D R L R Q I F M N L L S N A I S Y S L - - E G - G - - K UniRef90_A0YQV3_1128_1360 I A T P V P V I I N I D E L R I R Q V L I N L L S N A V K F T P - - E G - N - - H ê S17

18 UniRef90_E3PT03_185_421 S P I D N Q I Y L K S N P D L M K Q L I L N L I D N A I K Y S K - - E E - G - - F UniRef90_B7GGU1_348_587 T S N A P V C Y M Y G D L H R L K Q I F I N L I N N A I A Y T P - - A G - G - - R UniRef90_Q39S62_358_588 S G L S G A Q S V L A D P G R L E Q V L V N L L D N A I K Y T P - - E G - G - - A UniRef90_B8D0H3_363_592 D I K E K L P F V Y L I P E Q I E Q V L I N L V D N G I K Y T E - - P G - G - - R UniRef90_F5L948_225_461 M P P D E P A V V E A D P D R V Q Q I V I N L L S N A I T Y T P - - E G - G - - E UniRef90_E0NJE3_365_590 D V P M D I N S M Y V D R H G A D Q V F R N I F S N A L K Y T R - - K G - G - - E UniRef90_A7II27_682_911 E A P E T V P L L R A D P D R L T Q V V L N L I S N A V K F V P A - E G - G - - R UniRef90_B9DNB3_332_557 E I Q N H V M I D A E A S K I A Q V V T N L V S N A I N Y S S - - D E - S - - T UniRef90_A7HN49_181_412 K C E C K S T V Y G D F D R L V Q L V L N L V D N A V K Y T S I K E T - G E K K UniRef90_E6QIY0_358_593 D A A M H M V H A D Q E R I L Q V L A E L V S N A I K F S Q - - P H - T - - V UniRef90_F4L5H3_895_1123 S V S V D F P A A S A D E N R L Q Q I L Y N L L G N A V K F T E S - G - - Y UniRef90_A9AUY3_448_692 D I P E G I P P I F A D K D K V R Q I L T N L L S N A I K Y S P - - N G - G - - Q UniRef90_Q1IKU6_785_1018 H S T R G I L Y A D R D R M L Q T L T N L L S N A I K F S K - - P D - N - - T UniRef90_B5YD58_456_687 P E Y P T F I L G D N A R L A Q V F H N L L D N A I K Y S P - - N G - G - - N UniRef90_UPI00016BFB3D_364_592 S Y N K N K E Y L I E G D P A R I G Q V F H N I L S N A I K Y T E - - D N - G - - Q UniRef90_E8SIU9_335_566 Q I E P N V T A M A D E N K I S Q V I V N L M T N A V N Y S P - - E N - R - - T UniRef90_B9E797_336_561 E L E K N L F V L A E P S K L K Q V M I N L L S N A I N Y S P - - E D - A - - E UniRef90_Q8EPE4_222_453 E V P D Q L T M R A D K D R M K Q V L I N L I D N S I H Y T P - - S G - G - - D UniRef90_D5DL55_355_590 N G K V E D E F E A M I D S D R M E Q V L T N L I D N A I R H T D - - D Y - G - - E UniRef90_E0HX50_173_410 D V D E G I F V E G D F D R L V Q L L L N L V D N A V K Y T F A K E H - G P K E UniRef90_B8DZU7_455_686 P E Y P T F V L G D S A R L T Q V F H N L L D N A I K Y S P - - N G - G - - N UniRef90_D6GSX7_369_596 N L L G Q S R Q I W V D K T K M E Q V I I N I L S N A I K Y T E - - E N - G - - K UniRef90_D7CLL1_123_351 I F N R D L P P V K A D E D L V G Q V L I N L I D N A I K Y T S - - P G - G - - K UniRef90_Q6LXP6_406_633 E V E D I A I E A D K D R I T Q V L T N L I E N A I K F S P - - A N - E - - S UniRef90_Q67LR3_257_496 D V P E G L A R V M A D A D R L H Q V L D N L I S N A I K F S P - - Q G - S - - E UniRef90_A6LP23_177_414 V S P D K I E L I G D K D R L T Q M V L N L V D N A V K Y T S L K E K - G E K K UniRef90_B1HWE4_263_498 D I D G D V Q V M G D A N R I I Q I V T N L I T N A I T Y S P - - E N - T - - T UniRef90_Q2B2H9_351_585 S D Q S G N G W I K G D P H R L K Q V F I N L L T N A I T Y T P - - Q G - G - - K UniRef90_B8FA30_356_583 D C P D D L T A V F D P T M I E Q A V V N L L D N A V K Y S E - - E H - G - - V UniRef90_B5YJP0_360_619 E I Q P S L P F I N A D R E R L I Q V V I N L L S N A L K F T E K - G - - Y UniRef90_F0JJT0_240_468 E V E P G L A V R A D P A H V R Q V L T N L A G N G V K F S E - - R G - G - - T UniRef90_E1KHE7_585_832 V D N A D D T F G F I D K D L F E A V I A N L I S N A L K F T D - - S E - G - - Y UniRef90_C9LCS1_237_475 E S Y R P I V A E I D E V K L S L A L S N L V E N G I K Y N V - - E D - G - - W UniRef90_B7IEX9_176_421 V Y P D E L Y L V G D K D R L T Q M T L N L V D N A I K Y T S L K E K - G K K K UniRef90_Q8TMU8_474_720 E I D S N L S R V Y A D K E R I L Q V L S N L V T N A V K F S N - - E N - G - - C UniRef90_Q71Y67_354_593 E L E T P D L E Y S Y D P D R M E Q V L I N L I M N A I R H T G - - K E - G - - Y D G K V UniRef90_D5DMV7_352_585 D T L P E E A W I D G D S Q R L M Q V F V N L I G N A L M Y T M - - P G - G - - T UniRef90_A5D189_222_456 N L P E D L P V V Q G D P D M L S Q V L L N L I E N A V V Y T Q - - A G - G - - E UniRef90_C0Z7X8_357_585 P S P K P D I W L M T D K D C L Q Q I I L N L L T N A I A Y T Q - - E G - G - - K UniRef90_D4TMU3_804_1043 Q N S N S S S N S S S I Q L W A D P D R I I Q T L T N L F S N A I K F S E - - P G - S - - T UniRef90_Q0W6Q5_299_531 D V P L T L P M I S A D R S R I Q Q V L L N V L N N A I K Y T P - - D G - G - - Q UniRef90_B9E6M6_350_585 H Y S H H S K A W L D Y D K M I Q V I T N I I D N A L R Y T V - - A G - D - - V UniRef90_UPI000212C3B1_361_596 N V D G D F Y M E A D E D R L R Q I L I N L L S N G I S Y T P - - E G - G - - R UniRef90_A6UPU7_401_631 D I K D L L I N A D K D R I T Q V F T N L I E N A I K F S P - - A N - E - - S UniRef90_A5UQE0_447_687 D I P P H L P P V Y A D R D K V R Q V L V N L I T N A V K Y S P - - N G - G - - E UniRef90_A1HSJ7_400_654 A V D A G V P P Q V Q V D V R R F K Q I M Y N L L S N A V K F T P - - D G - G - - A UniRef90_F2JII5_342_577 G Y D K K E E Y F I E G D L S R I R Q I L H N I L S N A I K Y S P - - E P - G - - T UniRef90_Q8R6U6_339_575 N L S E T L Q K V N V D K D K V E Q M A M N I I N N A I K Y T P - - E G - G - - V UniRef90_Q8PT37_279_503 N V E P G F F V Y A D R T R F K Q I M Y N L V S N A V K F T Q - - K G - G - - S UniRef90_D8G5S6_939_1165 S S L S I S L W A D P D R I V Q T L I N L L S N A I K F S H - - T G - K - - I UniRef90_A8FJB5_372_608 N L P Q R E I Y V E I D Q D K I T Q V L D N I I S N A M K Y S P - - E G - G - - H UniRef90_A5ITL6_318_551 D I E K D V I V K A Q E S K I A Q V I T N L L T N A I N Y S Y - - E D - G - - D UniRef90_F6B3A9_217_450 V L P D K L P Q V P G D H G L L T Q V M V N L I D N A I K Y T P - - A S - G - - Q UniRef90_D9QVY1_356_592 D V P V D I T G I K G S R G Q L E R L Y I N L V D N G I K Y T S - - E G - G - - Q UniRef90_A9AA11_406_633 E I E E I A I D A D K D R I T Q V L T N L I E N A I K F S P - - A N - E - - S UniRef90_D5E8Z5_365_588 I N N S A I A K V W V D R V K F K Q I L H N L L S N A I K F T P - - E K - G - - E UniRef90_D2QCT5_672_903 C P T Q A Q P V V D G D R D R L M Q V L I N L L S N A I K F S P - - A G T G - - H UniRef90_A3DHV5_362_596 F T I G E E L E I V A D K D R M E Q V V L N I L T N A I K Y T P - - E G - G - - K UniRef90_B0ADS6_194_431 N F E D E D V C I Y G F R D N I K Q I F L N L I D N G I K Y T P - - K D - G - - H UniRef90_E7RC70_372_606 Q L P K E Q Y F V E I D P D K L T Q V I D N I I S N A L K Y S P - - E G - G - - K UniRef90_A0B593_141_369 R C E R A N T L I E C D P K K I M Q V L S N L L T N A I K Y T P - - D R - G - - Q UniRef90_D4IVS0_239_469 E S F R P V V A E V D E V K F T L V V T N L V E N A I K Y N D - - E G - G - - W UniRef90_B1YMQ3_615_835 V G P I M I D A D A E K I K Q L L N N L L S N A I K Y S P - - D G - G - - N UniRef90_C1F3B6_554_791 E A E P M D L W V D A D R I L Q T L G N L I S N A I K F S P - - A G - A - - D UniRef90_E7RJV0_231_462 E I Q P V T V L G D A N R L I Q V M M N L L I N A V T Y S S - - N Y - T - - E UniRef90_D3UPQ2_354_596 E L E T P N L E Y S Y D P D R M E Q V L I N L I M N A I R H T G - - K E - G - - Y Q G K V UniRef90_E0RD24_357_594 D I P D E L F M E A D E D K L Q Q I F L N L L S N G I N Y T L - - D G - G - - K UniRef90_F4LRR4_346_580 K L P E N I A H I L S N E D S L L Q I M I N L L D N A I K Y T P - - E G - G - - T UniRef90_Q7NDS1_97_ P G A L P E V W A D Q D R L E Q I L T N L I D N A L K Y S E - - H G - A - - P UniRef90_Q647M8_182_410 K L A E M P N I I G D A E R L R Q A I G N L L N N A I K F S E - - K T - G - - K UniRef90_B8G6G1_479_712 R T P L P P V F A D R D K V K Q I I V N L L S N A I K Y S P - - E G - T - - E UniRef90_B7GKK3_359_585 E M N D E L I V S F D P D R V E Q V L T N L I D N A L R H T D - - E G - G - - E UniRef90_C1DVW6_110_330 Q V P N K F D V Y A D E Q K L S I A L K N L I E N A I K Y N K - - Q N - G - - K UniRef90_C4L472_342_574 K H E G T V V L Q A D L N R M K Q V I M N L V A N A I N Y S P - - E G - S - - R UniRef90_D7GPR0_250_485 E S K R D V S A D I D E V K F S L A V N N L V E N A V K Y N K - - E D - G - - W UniRef90_B4S8R4_490_736 H V E P E T M Q V N A D P D A L K Q V V I N L A V N A I K F T P - - R D - G - - C UniRef90_A5GAH6_406_635 P S D L P R I C G D N K Q L R Q V F N N L V S N A V K Y S P - - K G - G - - A UniRef90_C6WZH9_504_741 K H F A D I E V F A D K E R L K Q V L T N L L T N A I K Y S P - - K A - N - - S UniRef90_B0TGE0_245_473 D V E T N L P L L P L S P D N L K Q V L V N L T E N A I K Y T P - - E G - G - - Q UniRef90_D5E978_387_612 N N K F D I D I N L D K L K F I Q I M S N L L S N A V K F T P - - D K - G - - K UniRef90_F5LRU7_361_595 N V D E A L F L E A D E D R L R Q I L I N L M A N G I N Y T P - - E G - G - - R UniRef90_Q4A159_364_598 E V P T E T I F T E I D P D K M T Q V F D N V I T N A M K Y S R - - G D - K - - R S18

19 G1 Box Input_pdb_SEQRES_2C2A - V - - K V I - - L D E - K D - - G G V L I I V E D N G I G I P D H A K UniRef90_A5IIT0_236_493 - V - - R V I - - L D E - K D - - G G V L I T V E D N G I G I P D H A K UniRef90_B9KAB5_220_473 - V - - K V V - - L D K - D E - - N G I L I V V E D N G I G I P E H A R UniRef90_UPI000210F96E_244_482 - V - - K I K - - L E R - L N - - D S V I I A V S D N G I G I P K E Y Q UniRef90_B7IDY9_224_462 - V - - N I S - - V E K - K E - - E N L S I I I E D N G I G I S Q E D K UniRef90_A6LN40_223_461 - V - - K I L - - V K K - L E - - D I L I I E V E D N G I G I P K S E Q UniRef90_A8F6Z9_244_486 - V - - K L S - - A E T - H Q - - D S V I I S V A D N G I G I E K Q H Q UniRef90_A7HK36_224_466 - V - - R I D - - V L D - E G - - E I V K I I V E D N G I G I P E S A F UniRef90_D5WTD2_380_614 - V - - D V R - - V L G T D P - - R E V K V E V A D T G V G I P P E D R UniRef90_Q67RF9_205_438 - I - - T L S - - A R R - D G - - D G V R I A V A D T G A G I P Q A D L UniRef90_B3E3A4_350_579 - V - - T V A - - A R L - E Q - - E R V V V A V S D T G I G I P A R D L UniRef90_C1P968_353_593 - I - - E I S - - L I E - R E - - N E L D L I V S D T G I G I S E K D L UniRef90_UPI000210FB4E_174_410 - V - - K V S - - A K L - Q D - - E S V V I K V E D T G P G I P K N A L UniRef90_D7AHN4_358_582 - I - - T L S - - A D E - A D - - G M V R V S V H D T G I G I P P K D L UniRef90_D5X969_353_585 - V - - V L S - - A T P - S G - - G G L K V S V K D T G I G I P P E S M UniRef90_D8PBL4_363_603 - I - - S M A - - I R N A T H M R A A T - P R - - P M I E I V V A D S G I G I P E A D R UniRef90_D7UVZ9_356_591 - V - - K V D - - V E E - S P - - H E V L F T V A D N G I G I P A K E I UniRef90_C5D651_348_580 - V - - E L M - - I E E - K E - - K E I L V H V K D T G I G I D E Q E I UniRef90_F4BLP5_336_567 - V - - T V T - - I R K - E N - - N Q A V I L V S D N G I G I P E D E Q UniRef90_Q3MGM2_1112_ V - - W L V - - A Q Q - Y G - - D E L L V T V K D N G R G I P A D K L UniRef90_B8G4S5_316_540 - I - - R I G - - A R A - E G - - E M A I V Y V A D Q G I G I P P E E Q UniRef90_E0FRK4_227_461 - V - - K V F - - L D I - K D - - K K A V I R I E D Q G Q G I P Q E E I UniRef90_B1ZT44_154_390 - V - - V V Q - - G R A T G A - - A M V E I A V C D D G P G I P A E A R UniRef90_C9RTX6_347_579 - V - - A V E - - A E E - T E - - K E V L I H V K D T G I G I E E K E I UniRef90_A4XMJ6_338_561 - I - - N V V - - L Q K - E E - - D K I K I I V K D N G P G I P E E D L UniRef90_UPI C5_286_519 - V - - K V F - - V K E - E R - - D E I T I K V Q D N G V G I A A S E L UniRef90_A5ILS8_180_410 - V - - W V R - - A Y D - T P - - D W V V L E V E D T G P G I P K E A Q UniRef90_B9K7H6_180_412 - V - - W V R - - A Y D - T P - - D W A V I E V E D T G P G I P K E A Q UniRef90_F2I7J7_353_596 - V - - W I S S T V T E - D D - - K Y V V V A I R D N G L G I P D E D I UniRef90_B5YHW7_385_611 - V - - K I T - - A E R - K E - - D M I E I V V E D T G I G I K K E D R UniRef90_C0A490_167_399 - I - - D I A - - I R P - K G - - S E I E I C V S D N G P G I P V A D L UniRef90_D6XVE0_357_597 - V - - K L R - - L E E - L P - - A G F R L D V E D T G A G I P E E D L UniRef90_A6CR52_338_570 - V - - S V S - - L K Q - S D - - S V I V I E V R D N G I G I E E S E F UniRef90_F5SLG1_353_585 - V - - I L S - - A G R - G E - - T D W W I R V A D T G I G I P G E D L UniRef90_A4FWN1_406_639 - I - - L V N - - G T L - E D - - E G L H L K V T D H G A G I P K K D M UniRef90_D3URW5_355_589 - V - - E V K - - L M E - R E - - S E V I I E V T D N G I G I P T K D I UniRef90_E8R548_187_428 - V - - K V V C R G D G - E A - - G M V A I D V I D N G L G I A S E E Q UniRef90_C9RD18_197_417 - V - - R I R - - G E Y - Q S - - G W V R I E V E D T G I G I P Q D C L UniRef90_C1PEW1_349_587 - V - - K V K - - E E E - A P - - G G V Y V H V E D N G S G I P D E D L UniRef90_Q1DCL6_267_509 - I - - T L Q - - V R G - S G - - K G V D L T V E D N G V G I A P Q E R UniRef90_A8F5W9_184_414 - V - - S I S - - V K K - E G - - S Q A V I R V S D T G P G M P R D A L UniRef90_Q2RFL3_239_485 - V - - T I R - - A A R - L G - - E L V Q V E V A D T G I G I P P E S L UniRef90_D1JAG2_243_467 - I - - S I K - - A I E - L N - - G Q V E V K V S D T G I G I P P E D L UniRef90_E5WJB7_351_584 - I - - Y I S - - A A K - T G - - S T V L T E I R D T G I G I E A S E I UniRef90_F6CIV3_222_456 - V - - T I R - - A M V - L E - - D Q L R V E V E D T G I G I P A E S L UniRef90_F6DKA0_218_449 - V - - T V G - - A S F - D G - - Q Q I R V F V K D T G I G I P E E S L UniRef90_E3Z246_18_252 - V - - E V K - - L I N - Q E - - A E V I I E V T D N G I G I P A K D I UniRef90_B5YHI6_153_383 - V - - K V R - - F F K - E N - - S K G V I S V E D T G I G I P K E H I UniRef90_A9GF59_268_496 - V - - T V R - - A R S - V D - - Q Q V T L A V A D D G P G I P P H H L UniRef90_E8WRA5_348_584 - I - - Q V E - - A A E - E G - - E M V R V S V R D N G I G I P E K D L UniRef90_Q097K6_265_506 - I - - A L S - - V R A - D R - - R W V A L S V E D N G V G I A P R D R UniRef90_E1KHP7_365_592 - I - - T V Y - - I G K - M Y - - S D A Y V K V V D S G I G I P E E D I UniRef90_A9AVU3_644_869 - I - - D V V - - V S N - G G - - D S V T L A V K D S G V G I A A D D Q UniRef90_A6UWZ1_410_640 - V - - T I E - - A F K - E N - - N M V H I T V K D N G P G I P K S E L UniRef90_D4Y401_348_580 - V - - E V I - - A E E - Q E - - E E T L V H V K D T G I G I E E A E I UniRef90_D7ATL0_341_566 - I - - K I M - - T E Y - D E - - S F A S L I V E D N G I G I P K E D L UniRef90_Q1PXZ9_292_526 - I - - T T N - - A A I - V D - - G K M Q V S V S D S G I G I K P E D R UniRef90_E7M6U2_368_603 - V - - S I G - - V E F V P S L D D N P - D N - - E R M R I R I S D T G I G I P E K D L UniRef90_D7BCF8_586_818 - V - - T L R - - A M L - E G - - D S L K I E V E D T G P G I P E S E R UniRef90_D0MGH9_109_352 - V - - E V R - - L Q E - Q D - - G S V R V A V V D N G I G I A P Q H I UniRef90_UPI000212A698_523_767 - V - - S V I - - I K Q - D D - - G G I V I S V S D T G I G I P L N M Q UniRef90_E1K386_410_648 - V - - E I Q - - A L K - E G - - N S V H I K I I D N G P G I P K K D L UniRef90_B9M7U2_251_475 - V - - T V G - - A E A - E G - - D Q V V V W V K D E G C G I P P E L Q UniRef90_Q3AAG9_206_440 - I - - E V T - - A A V - K G - - S W V E V V V K D T G I G I P E E S Q UniRef90_UPI0001E8932B_350_584 - V - - K V K - - V E E - V D - - K E V V I V V S D T G I G M E Q D E I UniRef90_C4G960_243_473 - V - - H V S - - L N A - D H - - Q Y C F V T V E D N G M G I P Q D S L UniRef90_C6J6K9_342_579 - V - - R V S - - A S I I G E G G - D E - - E K V R I L V S D T G I G I P K K D L UniRef90_A0YQV3_1128_ I L L K I E - - T E P - Q N - - N C V H L S V T D T G I G I A P E D Q ê S19

20 UniRef90_E3PT03_185_421 - V - - R L T - - L L E - D S - - K K V L F S V E D D G I G I P S E D I UniRef90_B7GGU1_348_587 - V - - T V Y - - V E K - D E - - K E L H V H V S D T G I G I E Q K E I UniRef90_Q39S62_358_588 - I - - S F S - - A A V - K E - - N M V R I G V K D T G V G I P P K D L UniRef90_B8D0H3_363_592 - V - - I L R - - A Y E - E N - - N R V V V E V E D N G I G I P E E D Q UniRef90_F5L948_225_461 - V - - R L S - - I D P W P D - - K G Y R I C V S D T G M G I N K E E I UniRef90_E0NJE3_365_590 - I - - K V T - - A K S - E G - - A S V E I I V E D N G I G I Q R E D L UniRef90_A7II27_682_911 - V - - H V S - - L H V - E G - - D G L V V R V K D N G P G V P E A E R UniRef90_B9DNB3_332_557 - V - - Y V R - - V Y Q - K D - - D K R I L E V E D H G I G I A P E E Q UniRef90_A7HN49_181_412 - V - - F V K - - C Y G - K E - - D K L V F E V Q D T G P G I P E D A Q UniRef90_E6QIY0_358_593 - V - - K L S - - A E S - A G - P G E V I I I V A D Q G R G I A A D K L UniRef90_F4L5H3_895_ I - - K I D - - A L E - 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I - - S L S - - L F V - S E - - G I L D I V I E D N G I G I P K E D V UniRef90_D7CLL1_123_351 - I - - V I R - - V K R - G G - - D Q V F T S I T D T G T G I P Q E S L UniRef90_Q6LXP6_406_633 - I - - L V S - - G V L - E D - - E H I H L K V T D R G A G I P K K D M UniRef90_Q67LR3_257_496 - V - - L I R - - A R E - T G - - D G V R F D V I D R G P G I P A D Q T UniRef90_A6LP23_177_414 - V - - L V E - - A Y K - D N - - N H V K L I V K D T G V G I P E K A Q UniRef90_B1HWE4_263_498 - V - - S I R - - L K E - N E - - T Y G I I E I E D Q G I G I E K H E I UniRef90_Q2B2H9_351_585 - I - - S V S - - L E E - K E - - D K V Q V E V K D T G M G I D Q D E I UniRef90_B8FA30_356_583 - V - - R V G - - A K A - E G - - G E V L I W V E D E G P G I S E E H L UniRef90_B5YJP0_360_619 - V - - K C K - - T A L - N A - - D E I V V S V E D S G I G I P E E E K UniRef90_F0JJT0_240_468 - V - - R L I - - A E R - S G - - R E V L F S V A D D G P G I P V D E Q UniRef90_E1KHE7_585_832 - I - - I I E - - L N K - L D - - T S F E I V V K D S G I G I P K D K L UniRef90_C9LCS1_237_475 - V - - H V T - - L N A - D H - - K Y F Y V S V E D S G I G I P Q E S I UniRef90_B7IEX9_176_421 - V - - T V E - - A W K - E N - - N S I K L V V K D T G V G I P E K A Q UniRef90_Q8TMU8_474_720 - V - - K V K - - A V Q - M D - - G F L K I T V A D D G I G I A A A D H UniRef90_Q71Y67_354_593 I L - - K Q T - - I D E - A R - - S N L V I T V S D N G S G I A E E D I UniRef90_D5DMV7_352_585 - V - - H V S - - L E E - E E - - E T I T V H V Q D T G I G I D S D E I UniRef90_A5D189_222_456 - V - - S I S - - A A A - T Q - - D E M K V D V K D N G I G I P P E S L UniRef90_C0Z7X8_357_585 - I - - S I K - - T E A - D S - - E N I T I Q V M D T G I G I P E K E L UniRef90_D4TMU3_804_ V - - Y L M - - T E L - Q K - - D Q V L V T V Q D T G R G I P E D K L UniRef90_Q0W6Q5_299_531 - I - - S I S - - V R D - E C - - D H L L I A V R D T G I G I P K E D I UniRef90_B9E6M6_350_585 - I - - S I H - - T D E - D D - - E N L I I R I S D T G V G I A P E H I UniRef90_UPI000212C3B1_361_596 - V - - R L K - - V E Q L V S G K E V T - E H - - D K V R F T I A D T G I G I P K K D L UniRef90_A6UPU7_401_631 - I - - M I I - - G K E T E N - - G D V H I T V K D N G A G I P K K D L UniRef90_A5UQE0_447_687 - I - - R L T - - I S - - D N V E L P P D H P - R G - - K F V R V A V S D Q G I G I A P E D L UniRef90_A1HSJ7_400_654 - I - - K V E - - C R D - A G - - E W L E I S V E D T G I G I A A T H A UniRef90_F2JII5_342_577 - L - - G V Y - - M K K - E N - - A Y V I V E I T D T G M G I P E E D L UniRef90_Q8R6U6_339_575 - I - - E I S - - T A Y - D E - - E G V T F T V K D N G I G I P K E D L UniRef90_Q8PT37_279_503 - V - - E V L - - G I V - S E - - K G V R V S V S D T G I G I S K D E I UniRef90_D8G5S6_939_ V - - W L T - - V Q Q - Q D - - R H L L F I V K D E G R G I P T E K L UniRef90_A8FJB5_372_608 - I - - T F T - - V D L - D E E K S L V L F S V K D E G I G I P K K D M UniRef90_A5ITL6_318_551 - I - - N V R - - V Y R - D D - - F R V I F E V Q D F G I G I K L E D Q UniRef90_F6B3A9_217_450 - V - - K V G - - V N L - N R - - E K V R V Y V A D T G I G I P Q E D L UniRef90_D9QVY1_356_592 - V - - K I K - - V Y E - D E - - D R V W S E I I D T G M G I P E E D L UniRef90_A9AA11_406_633 - I - - M I S - - A D L - E D - - E H V H L R V T D H G A G I P K K D M UniRef90_D5E8Z5_365_588 - V - - K I Y - - L S T - D D - - G T A Q I S V A D T G I G I P A E K I UniRef90_D2QCT5_672_903 - V - - T V C - - L T I - R D - - E R V N L T V R D N G I G I E P D V Q UniRef90_A3DHV5_362_596 - I - - T V Y - - I G R - M Y - - S E V Y V K V V D S G I G I P R E D L UniRef90_B0ADS6_194_431 - I - - E V V Q H Y D E - N R - - Q N I I L E F K D N G I G I P K E S L UniRef90_E7RC70_372_606 - V - - R F N - - M T V - E E - - G Y L L I Q I S D E G M G I P K E N V UniRef90_A0B593_141_369 - I - - E V V - - L G G - D A - - E N V L V S I R D N G I G I R E E D K UniRef90_D4IVS0_239_469 - V - - H I S - - L N S - D H - - Q F F Y I K V E D N G L G I P E N S V UniRef90_B1YMQ3_615_835 - V - - A I R - - I E N - H D - - G F A E I T I R D Q G I G I P Q E A M UniRef90_C1F3B6_554_791 - V - - L V R - - A S V - R G - K A E A V I E V R D H G R G I P P E K I UniRef90_E7RJV0_231_462 - I - - T I R - - L F R - K D - - N R A I I Q V E D Q G I G I E S S E I UniRef90_D3UPQ2_354_596 V L - - K Q T - - I D E - T N - - N K L I I T V S D N G S G I A E E D I UniRef90_E0RD24_357_594 - V - - K I K - - V L T I Q R E N - D T - - E K V V F T V S D T G I G I P K K D L UniRef90_F4LRR4_346_580 - I - - S I M - - V E E - T L - - E N V L I T V A D N G I G I A E D E L UniRef90_Q7NDS1_97_331 - V - - C V S - - A D L D P E - D R - - N V L W I A V R D L G I G I P E E N L UniRef90_Q647M8_182_410 - V - - I I E - - T K R - L G - - K N V Q F S I T D Y G I G I S K A D Q UniRef90_B8G6G1_479_712 - I - - Q L I - - V R Q A E A S D L P A G H P - A G - - Q W M L I T V Q D Q G I G I E P E D Q UniRef90_B7GKK3_359_585 - V - - R V I - - V D A - D E - - E V V R I S V Q D S G S G I P E E D L UniRef90_C1DVW6_110_330 - V - - I V K - - A V K - D S - - K Y T T I T V E D T G I G I P E E S I UniRef90_C4L472_342_574 - V - - E I A - - V E A - K A - - D S Y K L R V K D N G I G I A E K E V UniRef90_D7GPR0_250_485 - V - - R V T - - L D A - D H - - K F F Y L K V A D S G I G I P A E F K UniRef90_B4S8R4_490_736 - V - - R V S - - L S N - D A - - H W M V L T V K D S G V G I P E S D Y UniRef90_A5GAH6_406_635 - V - - R V A - - A R R - E K - - D C V V I T V R D E G I G I S P Q A V UniRef90_C6WZH9_504_741 - V - - N V E - - L W V - E D - - G R G I V S V E D F G I G M E A S E L UniRef90_B0TGE0_245_473 - V - - S V R - - A T R - G C - - D S V I L E V Q D T G I G I P E E S L UniRef90_D5E978_387_612 - V - - S V D - - I D F - V A - - E Y V Q V S V T D S G I G I P E H K L UniRef90_F5LRU7_361_595 - V - - S V A - - A E L A G S G T E - G - E D - - E R I R I S I K D T G I G I P K K D L UniRef90_Q4A159_364_598 - V - - E F H - - V K Q - N A L Y N R M T I R V K D N G I G I P I N K V S20

21 F Box G2 Box Input_pdb_SEQRES_2C2A D R I F E Q F Y R V D S S L T Y E V P - G T G L G L A I T K E I V E L H G G R I W V E - S E V G K G UniRef90_A5IIT0_236_493 D R I F E Q F Y R V D S S L T Y E V P - G T G L G L T I T K E I V E L H G G K I W V E - S E V G K G UniRef90_B9KAB5_220_473 E K I F E Q F Y R V D S S L T Y E V S - G T G L G L A I T K E I V E L H G G R I W V E - S E E G K G UniRef90_UPI000210F96E_244_482 S R I F E K F F R V Q S F K D Y K V E - G T G L G L T I C K E I V E L H G G K I W F E - S E P G K G UniRef90_B7IDY9_224_462 E K I F E K F Y R G D R S L T Y E V P - G T G L G L T I V Q E I I K L H G G K I N V N - S T L G E G UniRef90_A6LN40_223_461 E K I F E K F Y K I D R S L T Y E V P - G T G M G L A I V K E I V R L H G G N I E V E - S E E K K G UniRef90_A8F6Z9_244_486 N K I F E K F F R A D S V F D Y R T E - G T G L G L A I S K E I V E L H G G Q I W F E - S K P H E G UniRef90_A7HK36_224_466 D K I F E R F Y R V D N E L T Y A V P - G T G L G L A I V K E I V E L H E G N I L V E - S E V G K Y UniRef90_D5WTD2_380_614 D R V F E R F Y R V D K A R A R T T G - G T G L G L A I V K H V V Q L H G G R V G V E - S E V G K G UniRef90_Q67RF9_205_438 G R I F E R F Y R V D K A R S R A T G - G T G L G L A I A K H I V E A H G G T I G V E - S E V G K G UniRef90_B3E3A4_350_579 P R I F E R F Y R V D E G R S R E Q G - G T G L G L A I V K H I V Q L H G G E V Q V A - S E A G K G UniRef90_C1P968_353_593 P R I F E R F Y R V D K A R S R Q S G - G T G L G L A I V K H L V E S Y H G K I R V E - S E E G K G UniRef90_UPI000210FB4E_174_410 E R I F D R F Y R V D K G R S R K M G - G A G L G L S I V K T I V D R H N G K I Y V E - S E V G V G UniRef90_D7AHN4_358_582 P R I F E R F Y R V D A A R S R D E G - G T G L G L S I V K H I I Q L H G G N I T V E - S E H G K G UniRef90_D5X969_353_585 S R L F E R F Y R V D K A R S R E L G - G T G L G L A I V K H A L E A H G G T I K V E - S Q V G M G UniRef90_D8PBL4_363_603 P R V F E R F Y R V D K A R S R E L G - G T G L G L A I V K H I V E A H S G Q V W V E - G N T P R G UniRef90_D7UVZ9_356_591 E R V F E R F Y R V D K A R S R Y S G - G T G L G L S I V K H L T E Q L G G R I E V A - S V E G E G UniRef90_C5D651_348_580 P R I F E R F Y R V D K A R G R N S G - G T G L G L A I V K H L V E A H H G H I T V K - S T V G K G UniRef90_F4BLP5_336_567 D R I F E R F Y R V D K A R S R N S G - G T G L G L S I V K Y L V E N L N G S I A V E - S K L G L G UniRef90_Q3MGM2_1112_1344 D S I F E R F Q Q V D S S D S R N H D - G T G L G L A I C Q S I V Q Q H G G R I W A E - S V L S E G UniRef90_B8G4S5_316_540 D L I F E R F Y R V D N R L R R D R P - G S G L G L Y I T R A I V E A H G G R I W V E - S Q V G R G UniRef90_E0FRK4_227_461 K Y I F D K F Y R G K N N K Y S - G T G L G L A I S K K I I E E H K G T I T V E - S E V G K G UniRef90_B1ZT44_154_390 S R V F E R F Y R V D K A R S R E Q G - G T G L G L S I V K N L V Q A H G G E V R V E - S E L G R G UniRef90_C9RTX6_347_579 P R I F E R F Y R V D K A R S R D S G - G T G L G L A I V K H L V E A H H G Y I T V A - S K V G R G UniRef90_A4XMJ6_338_561 P R I F E R F Y R V D K A R S R E L G - G S G L G L S I A D E I V K A H G G K I L V E - S K V G S G UniRef90_UPI C5_286_519 P R I F E R F Y R I D K A R S R N S G - G T G L G L A I V K H I V E A H H G E I D V E - S S E G V G UniRef90_A5ILS8_180_410 S R I F E K F Y R V D K A R S R K M G - G T G L G L T I V K T I V D K H G G R I E V E - S E I N Q G UniRef90_B9K7H6_180_412 S R I F E K F F R V D K A R S R K M G - G T G L G L T I V K T I V D R H G G K I E V E - S E V G Q G UniRef90_F2I7J7_353_596 P R I F E R F Y R V D K T R S T A S G - G T G L G L S I V R N L V A S M S G K I D V V - S E L N E G UniRef90_B5YHW7_385_611 D K L F Q P F S Q L E T T Y T K K Y Q - G T G L G L A L S K S L V E L H G G K I W C E - S E Y G K G UniRef90_C0A490_167_399 P H I F E R F Y R V D K G R S R E T G - G T G L G L S I V K H I V Q L H G G R V W V E - S E P G R G UniRef90_D6XVE0_357_597 P F V F E R F Y K A D K A R T R G R A - G T G L G L A I V K N I V E A H K G R V S A H - S R M G E G UniRef90_A6CR52_338_570 P R I F E R F Y R V D K A R S R D S G - G T G L G L A I V K H L I E A H G G T I G V E - S K V N E G UniRef90_F5SLG1_353_585 P R I F E R F Y R V D K A R S R E S G - G T G L G L A I V K H L V E A H Q G E I Q V T - S R A G K G UniRef90_A4FWN1_406_639 E K V F D R F Y Q V D S S T K R K K G - G S G L G L A V C K S I V E A H G G S I W V E - S E L G K G UniRef90_D3URW5_355_589 D R V F E R F Y R V D K A R S R H S G - G T G L G L S I V K H L V E N C G G R I E V E - S Q E E V G UniRef90_E8R548_187_428 T R I F E R F Y R V D K A R S R E R G - G T G L G L A I V K H L V Q A L R G E I E L Q - S R V G A G UniRef90_C9RD18_197_417 P R V F E R F F R V D R A R S R A S G - G T G L G L S I V K H I V E L H G G K V G V E - S E L G K G UniRef90_C1PEW1_349_587 P F V F E R F Y K A D K A R T R G K S - G T G L G L A I A K N I V E G H G G R I F V K - S K L G E G UniRef90_Q1DCL6_267_509 T R I F E R F Y R V D N L L T R R T E - G S G L G L A I T R R I I E T H G G R I S V Q - S E P G K G UniRef90_A8F5W9_184_414 N R I F D R F Y R V D K G R S R K M G - G S G L G L S I V K T I V D R H N G Q I F V E - S E P G A G UniRef90_Q2RFL3_239_485 P R V F E R F Y R V D K A R S R E M G - G T G L G L A I V K H I V E S H G G S I S V T - S R P G Q G UniRef90_D1JAG2_243_467 E K V F D K F Y Q V D S T L T R E A G - G T G L G L A I C K G I I E A H N G H I W A E - S E L G K G UniRef90_E5WJB7_351_584 P R I F E R F Y R V D K A R S R N S G - G T G L G L A I V K H L V E A H K G S I S V K - S E V G K G UniRef90_F6CIV3_222_456 P R I F E R F Y R V D K A R S R E L G - G F G I G L A I V K H I I R A H G G K I E V E - S T P G K G UniRef90_F6DKA0_218_449 S R V F E R F Y R V D K A R S R D V G - G T G L G L S I S K H I V E A H G G K I W A E - S H - S E G UniRef90_E3Z246_18_252 D R V F E R F Y R V D K A R S R H S G - G T G L G L S I V K H L V E N C G G R I E V E - S Q E E V G UniRef90_B5YHI6_153_383 H R I G E R F Y R V D K A R S R Q L G - G T G L G L A I V K H L V L A H G W Q L Q I E - S E V E K G UniRef90_A9GF59_268_496 G R I F E R F Y R V D A G R S R D L G - G T G L G L A I V K H L V E L M N G S I E V E - S A I G R G UniRef90_E8WRA5_348_584 P R L F E R F Y R V D E A R S R E R G - G T G L G L S I V K H I V M A H G G S V F V E - S T V G K G UniRef90_Q097K6_265_506 Q R I F E R F Y R V D N L L T R K T E - G S G L G L A I T K R I I E A H G G R I S V Q - S K L G K G UniRef90_E1KHP7_365_592 K R V F E R F Y R V D K A R S R E M G - G T G L G L S I A K E I I E A H K G S I S I S - S Q L G K G UniRef90_A9AVU3_644_869 K H I F T R F F R S E N P L K E Q A G - G T G L G L N I T K S L V E L H G G K I W F D - S E E G R G UniRef90_A6UWZ1_410_640 T K I F D I F Y Q V N S S A K R I K S - G S G L G L A I C K S I V E S H G G K I W V E - S K F G K E UniRef90_D4Y401_348_580 P R I F E R F Y R V D K A R S R H S G - G T G L G L A I V K H L V E A H H G H I T V K - S A V G K G UniRef90_D7ATL0_341_566 P R I F E R F Y R V D K A R S R E L G - G T G L G L S I V K Q I V E L H K G E V N I E - S E V G K G UniRef90_Q1PXZ9_292_526 E K V F K E F W Q A D S S F S R K Y E - G T G L G L A L T R R I V E M H G G K I W L K - S E Y G K G UniRef90_E7M6U2_368_603 P R I F E R F Y R V D K A R S R S S G - G T G L G L S I V K H L T E L H H G T I S V E - S E A G K G UniRef90_D7BCF8_586_818 S R L F Q R Y G R T Q S A V S R G V S - G T G L G L Y I S K A I V E A H G G R I W V E - S E V G K G UniRef90_D0MGH9_109_352 P R L T E R F Y R V D R S R S R E Q G - G T G L G L A I V K H I L N A H Q T R L E I E - S T P G K G UniRef90_UPI000212A698_523_767 E N I F D L F T Q V D G S I K R K Y D - G T G L G L A I V K Q Y V E M H N G K V W V E - S E E D K G UniRef90_E1K386_410_648 D R I F D R F Y Q V D S P E K R I K G - G S G L G L A V C K S I I E T H G G T I W V E - S K L G S G UniRef90_B9M7U2_251_475 E K I F E K F Y R I D N T D R R H I G - G T G L G L A L V R E I V A A H G G R V W V T - S E V G R G UniRef90_Q3AAG9_206_440 K R I F E R F Y R V D K A R S R E L G - G T G L G L A I V K H I I E L H N G K V W V K - S K V G E G UniRef90_UPI0001E8932B_350_584 P R I F E R F Y R V D K A R S R N S G - G T G L G L A I V K H I V E A H H G S I S V T - S E L N K G UniRef90_C4G960_243_473 D R I F E R F Y R V D K S H S R E I G - G T G L G L A I T Q N A I R M H H G E I R V A - S E L G Q G UniRef90_C6J6K9_342_579 P R I F E R F Y R V D K A R S R S S G - G T G L G L S I V K H L V D L H R G T I R V E - S K V G E G UniRef90_A0YQV3_1128_1360 V R L F E P F V Q I D S S L S R R Y S - G T G L G L A L V R Q I T Q M H A G T V S L D - S E V G R G ê G3 _ S21

22 UniRef90_E3PT03_185_421 D R I F E R F Y R V D K A R S K K V G - G T G L G L A I V K H I V L N L N G N I K V H - S E L N K G UniRef90_B7GGU1_348_587 P R I F E R F Y R V D K A R S R N S G - G T G L G L A I V K H L V E A H H G T I S V K - S E V G I G UniRef90_Q39S62_358_588 P R I F E R F Y R V D T A R S R D E G - G T G L G L S I V K H I V Q L H G G A V G V E - S E P G K G UniRef90_B8D0H3_363_592 G R I F E R F Y R V D K A R S R S M G - G T G I G L S I V K H I I K N H D S E I K V E - S E P G K G UniRef90_F5L948_225_461 P R I F E R F Y R V D R A R S R A S G - G T G L G L A I V K H L V E A H R G E I T V E - S E P G K G UniRef90_E0NJE3_365_590 N K I F E R F Y R V E K S R S R E M G - G T G L G L S I A K E I L E S M G G R I S I E - S E I G V G UniRef90_A7II27_682_911 D T I F E K F R Q G G D A L T R P P - - G T G L G L P I S R R I V D H F G G R M W L E - N Q D G S G UniRef90_B9DNB3_332_557 K H V F E R F Y R V D K A R S R D S G - G T G L G L S I T K H I V E A Y Q G R I Q I E - S E P D V G UniRef90_A7HN49_181_412 K R L F E R F Y R V D K A R S R K V G - G T G L G L S I V K M I A D K H N A T I S F E - S K V G E G UniRef90_E6QIY0_358_593 E V I F E R F H Q G D A S D A R A L G - G T G M G L A L C R Q I V R Q H G G R I W A E - S E P D K G UniRef90_F4L5H3_895_1123 D A I F Q E F T Q G D A S T T R A F S - G T G L G L S I S K R L V E L H G G T M W L E - S E I D Q G UniRef90_A9AUY3_448_692 P K L F T R F F R V D N S T T R K I G - G T G L G L S I T K A L I E L H G G R I W A T - S T L G R G UniRef90_Q1IKU6_785_1018 Q T I F E R F Q Q V D A S D S R D K G - G T G L G L A I C R S I V Q Q H G G S I W V D - S I D G K G UniRef90_B5YD58_456_687 P H I F D R F Y R V D S S L R K S T S - G T G L G L S I V K S I I E A H G G K I S V A - S K V G E G UniRef90_UPI00016BFB3D_364_592 E R I F E R F Y R A D K A R S R K M G - G T G L G L S I A K E M V E L H G G S I K M E - S A L G H G UniRef90_E8SIU9_335_566 Y R I F E R F Y R V D K A R S R D S G - G T G L G L S I T K H I I E A Y Q G N I E V A - S E L G K G UniRef90_B9E797_336_561 T R I F E R F Y R V D K A R S R D S G - G T G L G L A I V K H I I E V F N G E I D V E - S E L G K G UniRef90_Q8EPE4_222_453 T R V F E R F Y R V D K A R S R N T G - G T G L G L A I V K H I V E V H K G K I D V E - S T L N Q G UniRef90_D5DL55_355_590 P F V F E R F Y K A D K A R T R G K S - G T G L G L A I A K N I V E A H Q G I I A V E - S E L N E G UniRef90_E0HX50_173_410 K H I F E R F Y R V D K A R S R K M G - G T G L G L A I T R F I V E K H G G T I S L E - S E Y G T G UniRef90_B8DZU7_455_686 P H I F D R F Y R V D S S L R K S T S - G T G L G L S I V K S I V E A H G G K I S A T - S K V G E G UniRef90_D6GSX7_369_596 E H I F D R F Y R V D K G R S R Q Q G - G T G L G L S I A K H I V E E H G G S I S V R - S D F G K G UniRef90_D7CLL1_123_351 P R L F E R F Y R V D K A R S R E L G - G T G L G L A I V K H I V E S H G G E V F V E - S E L G K G UniRef90_Q6LXP6_406_633 E K I F N R F Y Q V D S S T K R K K G - G S G L G L A V C K S I V E A H K G S I W V E - S E L G K G UniRef90_Q67LR3_257_496 G L I F E R F Y R V D N A A S R K T G - G T G L G L T I C K A I V E E H G G Q I W V E - S A L G E G UniRef90_A6LP23_177_414 K K L F E R F Y R V D K A R S R K M G - G T G L G L S I V K T I V E K H N G T I E F T - S K E G V G UniRef90_B1HWE4_263_498 A R V F E R F Y R V D R A R S R N S G - G T G L G L A I V K H L V E A H H G R I Q V E - S E V G V G UniRef90_Q2B2H9_351_585 P R I F E R F Y R I D K A R S R N S G - G T G L G L A I V K H L V E A H K G S I T V E - S E V G K G UniRef90_B8FA30_356_583 P R L F E R F Y R V D K A R S R N L G - G T G L G L A I V K H I M A A H G G H V S V K - S E K G K G UniRef90_B5YJP0_360_619 E R I F E K F K Q V G D L I R G K P K - G T G L G L A I S K Q I V E A H G G K I W V E - S E L G K G UniRef90_F0JJT0_240_468 E R I F L K Y Y R - E P G V R D S I D - G A G L G L A I A R R I V L A H D G R I W V E - S E P G R G UniRef90_E1KHE7_585_832 D T I F D R F S Q I E N S K S S K V R - G T G I G L A Y T K E I V E L H E G K I S V S - S I L G K G UniRef90_C9LCS1_237_475 D R I F E R F Y R V D K S H S R E I G - G T G L G L A I T R N A I V M H R G A I K V K - S E E H K G UniRef90_B7IEX9_176_421 K K L F E R F Y R V D K A R S R K M G - G T G L G L S I V K T I V E K H N G V I E F T - S K E G V G UniRef90_Q8TMU8_474_720 E K L F K P F S Q I D S S F S K R Y Q - G T G L G L A L V K E I V Q L H G G T V W F E - S E V G K G UniRef90_Q71Y67_354_593 P Y L F E R F Y K V D K A R K R G K A V G T G I G L A I V K N I V E A H N G K I S V E - S E L G K G UniRef90_D5DMV7_352_585 S R I F E R F Y R V D K A R S R N S G - G T G L G L A I V K H L V E A H K G E I E V K - S E V N K G UniRef90_A5D189_222_456 S R V F E R F Y R V D K A R S R E Q G - G T G L G L S I V K H I I D A H R G S V Q V E - S K V G I G UniRef90_C0Z7X8_357_585 T R I F E R F Y R V D K A R S R D S G - G T G L G L A I V K H L V E N L H G H I S V E - S K E G R G UniRef90_D4TMU3_804_1043 E S I F E R F Q Q V D S S D S R N H D - G T G L G L A I C K S I V Q Q H G G K I W V K - S V L G Q G UniRef90_Q0W6Q5_299_531 E N I F S G F Y H S G Y K L S Y E Y K - G A G L G L A I S R K I V E S H G G K I W A D - S E P G K G UniRef90_B9E6M6_350_585 D N I F E R F Y K V D Q A R T R G K H - G T G L G L F I V K S I V E G H H G S I N V D - S T V G K G UniRef90_UPI000212C3B1_361_596 P R I F E R F Y R V D K A R S R S S G - G T G L G L S I V K H L V E L H K G T I R V E - S E V G V G UniRef90_A6UPU7_401_631 E K I F D Q F Y Q V D S S T K R K K G - G S G L G L A V C K S I I Q A H G G T I W V E - S E L G R G UniRef90_A5UQE0_447_687 P R I W E R F Y R V D N G N T R R I G - G T G L G L S I A K A L V E L H G G R I W A E - S K L N K G UniRef90_A1HSJ7_400_654 E H I F E P F Y Q V S G N L T A K T P - G T G L G L A I T R Q L V E L H G G R I W L E R S E P G Q G UniRef90_F2JII5_342_577 E R I F E R F Y R V D K A R S R K M G - G T G L G L S I A K E L M L L H G G D I K I E - S Q L G Q G UniRef90_Q8R6U6_339_575 P R I F E R F Y R V D K A R S R E L G - G T G L G L S I V K Q I V E L H K G K V K I E - S E L G K G UniRef90_Q8PT37_279_503 K Q L F K P F K Q I D S T L S R K Y E - G T G L G L V L S K K F V E M H G G R I W V E - S E P G K G UniRef90_D8G5S6_939_1165 E S I F E R F Q Q V D S S D S R N H E - G T G L G L A I C R S I V Q Q H D G H I W V E - S V L G E G UniRef90_A8FJB5_372_608 D K I F E R F Y R V D K A R T R K L G - G T G L G L A I A K E M V Q A H G G D I W A D - S I E G K G UniRef90_A5ITL6_318_551 Q R I F E R F Y R V D K A R S R D S G - G T G L G L S I T K H I V E A H Q G N I E V N - S Q V G K G UniRef90_F6B3A9_217_450 P R I F E R F Y R V D K A R S R V M G - G T G L G L A I C K H I V E V H G G Q I E V E - S G P G - G UniRef90_D9QVY1_356_592 P R I F E R F Y R V D K T R S R K L G - G T G L G L S I V K H I L E R H N G G I E V E - S K V E E G UniRef90_A9AA11_406_633 E K V F N R F Y Q V D S S T K R K K G - G S G L G L A V C K S I V E A H K G S I W V E - S E L G K G UniRef90_D5E8Z5_365_588 E D I F D P F K Q V N S S S S R E Y E - G T G L G L A L V K K Y V E M H D G E I W V E - S E V G A G UniRef90_D2QCT5_672_903 Q L I F E K F Y Q A T N Q T L R K P K - G S G L G L A I T K K I I A L H N G Q I T V S - S V P G E G UniRef90_A3DHV5_362_596 G R I F E R F Y R T D K A R S R E M G - G T G L G L A I A K E I V E A H K G S I S V A - S E P G K G UniRef90_B0ADS6_194_431 N R I F E R F Y R V D K A R S R D I G - G T G L G L A I T K H M V K S L G G N I T V E - S I L G I G UniRef90_E7RC70_372_606 D R I F D R F Y R V D R A R S R E M G - G T G L G L A I S K E M I N A H G G V I W A E - S K L G K G UniRef90_A0B593_141_369 D R I F E K F F V M S N E K S A D I - - G M G L G L S I T K D I V E A H G G R I W F E - S R F G E G UniRef90_D4IVS0_239_469 D H I F E R F Y R A D K S H S R E I G - G T G L G L A I T K S A I A M H N G E I K V R - S K L G E G UniRef90_B1YMQ3_615_835 S K L F D K F Y R V D N S E T R K I G - G T G L G L S I C K E I V K H H N G T I D V E - S T I G A G UniRef90_C1F3B6_554_791 S L I F E R F Q Q V D A S D S R A M G - G T G L G L A I C R R I V R Q H G G Q I W V E - S E V G K G UniRef90_E7RJV0_231_462 G R L F E R F Y R V D R A R S R N S G - G T G L G L S I V K H L I E A H H G H V E V D - S T V G V G UniRef90_D3UPQ2_354_596 P Y L F E R F Y K A D K A R K R G K A V G T G I G L A I V K N I V E A H G G K I S V K - S E L G K G UniRef90_E0RD24_357_594 P R I F E R F Y R V D K G R S R N S G - G T G L G L S I V K H L V D L H H G T L S V E - S E L G L G UniRef90_F4LRR4_346_580 D R I F E R F Y R V D R A R S G E I G - G T G L G L A M V K H L V K G L E G W I K V E - S E L E K G UniRef90_Q7NDS1_97_331 D Q I F N R F S R I D S P L T R E R E - G T G L G L Y I T K S L V E S L G G T I H V E - S R Y G V G UniRef90_Q647M8_182_410 K K L F K P F T Q I D S S M G R E H V - G T G L G L A I T K G I I Q A H N G K I W A E - S E L G K G UniRef90_B8G6G1_479_712 A R L F E R F H R V D N S N T R Q K S - G V G L G L Y I T R L L V E L H G G R I W V Q - S T V G V G UniRef90_B7GKK3_359_585 P F V F E R F Y K A D K A R T R G R S - G T G L G L A I A K N I V E A H K G T I T V H - S K L G E G UniRef90_C1DVW6_110_330 P L I F E R F Y R V D K S R S R N I G - G T G L G L S I V K H I T E A H K G K V W V E - S K L G E G UniRef90_C4L472_342_574 S R I F E R F Y R V D K A R S R N S G - G T G L G L A I V K H I I D L H H A T I D V E - S E E G K G UniRef90_D7GPR0_250_485 E R V F E R F Y R V D K A R S R E T G - G T G L G L A I T K S V V L M H H G A I R V D - S V E G E G UniRef90_B4S8R4_490_736 D K I F E K F Y R V E R P - G E E I E - G T G L G L P I V K E I I A A H K G S I E V N - S K K D F G UniRef90_A5GAH6_406_635 E R I F E R F Y R V N N T A C R V F G - G T G L G L A L V R E I V T A H G G E V W V E - S T L G K G UniRef90_C6WZH9_504_741 G K I F E R F Y R V S G D D E K T F P - G F G I G L Y I V K D I I Q R L D G K I W V E - S E K N Q G UniRef90_B0TGE0_245_473 P R I F E R F Y R V D K A R S R E L G - G T G L G L A I V K H I I E R S G G R V T V Q - S K V G V G UniRef90_D5E978_387_612 N M I F D P F I Q A D S S N T R K Y G - G T G L G L A L V K E Y V E M H G G T I W V E - S E L G K G UniRef90_F5LRU7_361_595 P R I F E R F Y R V D K A R S R S S G - G T G L G L S I V K H L V E M H H G T I R V E - S E V G Y G UniRef90_Q4A159_364_598 D K I F D R F Y R V D K A R T R K M G - G T G L G L A I S K E I V E A H N G R I W A N - S V E G Q G S22

23 Input_pdb_SEQRES_2C2A S R F F V W I P K D R A G E D N R Q D N - UniRef90_A5IIT0_236_493 S R F F V W I P K D R A G E D N R Q D N - UniRef90_B9KAB5_220_473 S R F F V W I P Y D H G T E D N R UniRef90_UPI000210F96E_244_482 S I F Y V UniRef90_B7IDY9_224_462 T T F E L V I P UniRef90_A6LN40_223_461 S I F R V N I P UniRef90_A8F6Z9_244_486 T V F Y V K I P Q UniRef90_A7HK36_224_466 T K F I V T L P K N UniRef90_D5WTD2_380_614 S R F F V Q F P R UniRef90_Q67RF9_205_438 S T F T V I L P UniRef90_B3E3A4_350_579 S T F T V T L P UniRef90_C1P968_353_593 S T F I V T L P R T Q T R P D UniRef90_UPI000210FB4E_174_410 T T F T V V L P UniRef90_D7AHN4_358_582 T T F UniRef90_D5X969_353_585 S K F T F Y L P UniRef90_D8PBL4_363_603 S R F V V H L P UniRef90_D7UVZ9_356_591 T T F S I H L P K T K UniRef90_C5D651_348_580 T T F T V H F P K UniRef90_F4BLP5_336_567 T T F I V R L P UniRef90_Q3MGM2_1112_1344 S N F Y F T L P UniRef90_B8G4S5_316_540 S R F L F T L P L S R UniRef90_E0FRK4_227_461 S V F T I E L P L K D R UniRef90_B1ZT44_154_390 A R F F F T L P E A E A UniRef90_C9RTX6_347_579 T V F T I H F P K UniRef90_A4XMJ6_338_561 T T F T V V L P UniRef90_UPI C5_286_519 T T F I V K V S K E UniRef90_A5ILS8_180_410 T V M R V F L P K UniRef90_B9K7H6_180_412 T L M R V Y L P K G R UniRef90_F2I7J7_353_596 S T F T V Y L P K N UniRef90_B5YHW7_385_611 S R F G F S F P UniRef90_C0A490_167_399 A S F H F T L P UniRef90_D6XVE0_357_597 T T F S V F L P F S Q G E D UniRef90_A6CR52_338_570 T T F T V T L N K UniRef90_F5SLG1_353_585 T E I T L T F P R UniRef90_A4FWN1_406_639 S T F H I I L P I S V E G Q UniRef90_D3URW5_355_589 S T F R V T L P K UniRef90_E8R548_187_428 S R F T V R L P R Q R UniRef90_C9RD18_197_417 S R F W V Y L P S P Q UniRef90_C1PEW1_349_587 T T F S F F L P K UniRef90_Q1DCL6_267_509 S R F T I H L P A G K A UniRef90_A8F5W9_184_414 T S F T V K L P V E R UniRef90_Q2RFL3_239_485 S H F F F T L P I A A E E G G R S N Y Q E UniRef90_D1JAG2_243_467 S T F UniRef90_E5WJB7_351_584 T S F I I E L P K UniRef90_F6CIV3_222_456 S L F Y F T L P UniRef90_F6DKA0_218_449 S T F I F T L P UniRef90_E3Z246_18_252 S T F R V T L P K UniRef90_B5YHI6_153_383 T K V K I V I P UniRef90_A9GF59_268_496 T Q F T V R L A R UniRef90_E8WRA5_348_584 S V F S F T L P R A G D UniRef90_Q097K6_265_506 S L F T L Q L P A G K A UniRef90_E1KHP7_365_592 T E V I V R L P UniRef90_A9AVU3_644_869 T T F N V Q L P UniRef90_A6UWZ1_410_640 S T F H I L L P L D K UniRef90_D4Y401_348_580 T T F T V H F P K UniRef90_D7ATL0_341_566 T I V R V K L P UniRef90_Q1PXZ9_292_526 S T F Y F T I P UniRef90_E7M6U2_368_603 T T F N I E L P UniRef90_D7BCF8_586_818 T T F S L T L P UniRef90_D0MGH9_109_352 S T F A F R L P V A L A G R S UniRef90_UPI000212A698_523_767 S T F T F T I D T K R N Q E UniRef90_E1K386_410_648 S V F H I I I P UniRef90_B9M7U2_251_475 S T F F I S L P UniRef90_Q3AAG9_206_440 S A F G F T I P UniRef90_UPI0001E8932B_350_584 T A F T V K L S K E UniRef90_C4G960_243_473 T T F D V R I P UniRef90_C6J6K9_342_579 T T F I L E L P UniRef90_A0YQV3_1128_1360 S T F I V T L P S23

24 UniRef90_E3PT03_185_421 S K F I I E F P K N UniRef90_B7GGU1_348_587 T T F T V H F R A D A C A D E UniRef90_Q39S62_358_588 S E F W F T I K K UniRef90_B8D0H3_363_592 S L F R F Y L N K UniRef90_F5L948_225_461 S I F C V Y L Y Q K R UniRef90_E0NJE3_365_590 T K M T L R F P UniRef90_A7II27_682_911 A C F A F R L P UniRef90_B9DNB3_332_557 T T F UniRef90_A7HN49_181_412 T T F R V Y F K K UniRef90_E6QIY0_358_593 S R F L F T L P A G S T A I D S UniRef90_F4L5H3_895_1123 S K F F F S L Q K UniRef90_A9AUY3_448_692 T T F W V T L P UniRef90_Q1IKU6_785_1018 S E F F I L L P R UniRef90_B5YD58_456_687 T T F T I R L P R UniRef90_UPI00016BFB3D_364_592 T UniRef90_E8SIU9_335_566 S R F K V V L P E UniRef90_B9E797_336_561 S T F UniRef90_Q8EPE4_222_453 T T I H I Y I P K UniRef90_D5DL55_355_590 T T F F I K L P K UniRef90_E0HX50_173_410 T I L K V K L P UniRef90_B8DZU7_455_686 T T F T I R L P R UniRef90_D6GSX7_369_596 T V F T I S L P UniRef90_D7CLL1_123_351 S T F G F S L W V UniRef90_Q6LXP6_406_633 S T F H I I L P UniRef90_Q67LR3_257_496 S T F S F T I P V E P G G UniRef90_A6LP23_177_414 T K F I V T L P UniRef90_B1HWE4_263_498 T T M I V M I P K N UniRef90_Q2B2H9_351_585 T T F K V Q L H R E UniRef90_B8FA30_356_583 A A F UniRef90_B5YJP0_360_619 S T F Y F T I P L K R K E E F N E N I N - UniRef90_F0JJT0_240_468 A T F R F T L P UniRef90_E1KHE7_585_832 S V F A V S L P UniRef90_C9LCS1_237_475 T T F S V R I P L N Y I G UniRef90_B7IEX9_176_421 T K F V V T L P I N R E E D N N UniRef90_Q8TMU8_474_720 S V F G F S I P UniRef90_Q71Y67_354_593 S D F I I T L P UniRef90_D5DMV7_352_585 T T F S V K L H K UniRef90_A5D189_222_456 S T F S F V I P UniRef90_C0Z7X8_357_585 T T F T V T L P UniRef90_D4TMU3_804_1043 S S F Y F T L P UniRef90_Q0W6Q5_299_531 S T F F I S L P K UniRef90_B9E6M6_350_585 T T F V I S I P K V D D T K N N K UniRef90_UPI000212C3B1_361_596 T K F I I E M P UniRef90_A6UPU7_401_631 S T F H I V L P UniRef90_A5UQE0_447_687 S T F Y F T L P UniRef90_A1HSJ7_400_654 S R F T F V L P V R Q D G UniRef90_F2JII5_342_577 T K V I L N F P E UniRef90_Q8R6U6_339_575 T V V T V Q L P Y S K UniRef90_Q8PT37_279_503 S T F T F E V P UniRef90_D8G5S6_939_1165 S I F Y F M I P UniRef90_A8FJB5_372_608 T T V T F T L P Y N E E Q E D D UniRef90_A5ITL6_318_551 S T F K V I L K D UniRef90_F6B3A9_217_450 S T F S F T L P UniRef90_D9QVY1_356_592 T K F I F W L P K P K UniRef90_A9AA11_406_633 S T F H I I L P UniRef90_D5E8Z5_365_588 S T F UniRef90_D2QCT5_672_903 A E F T C S L P UniRef90_A3DHV5_362_596 T E V T V K L P UniRef90_B0ADS6_194_431 S D F I V T I P V D UniRef90_E7RC70_372_606 T S I F F T L P F E E D D G G E UniRef90_A0B593_141_369 S T F Y F T L P UniRef90_D4IVS0_239_469 T T F D V R I P UniRef90_B1YMQ3_615_835 S T F T I R F P UniRef90_C1F3B6_554_791 S S F F F T V A R N V A UniRef90_E7RJV0_231_462 T T F T I Y L P UniRef90_D3UPQ2_354_596 S D F I I T L P L E K UniRef90_E0RD24_357_594 T T F T V E L P UniRef90_F4LRR4_346_580 T S F T V Y I P K UniRef90_Q7NDS1_97_331 S T F T V S L P A V Q A G UniRef90_Q647M8_182_410 S T F Y F V I P UniRef90_B8G6G1_479_712 S S F S F T L P UniRef90_B7GKK3_359_585 T T F T F T L P UniRef90_C1DVW6_110_330 S K F F I K I P UniRef90_C4L472_342_574 T T F T I T F P R UniRef90_D7GPR0_250_485 S T F M V R I P UniRef90_B4S8R4_490_736 S T F F V R L P L S T S G UniRef90_A5GAH6_406_635 S T F F V S L P UniRef90_C6WZH9_504_741 S K F Y F S L P UniRef90_B0TGE0_245_473 S T F T V T F P UniRef90_D5E978_387_612 S T F T F T I P UniRef90_F5LRU7_361_595 T E F I I E L P UniRef90_Q4A159_364_598 T S I F I T L P S24

25 Fig. S6 Competition of HK853 activity in the presence of guanidine derivatives 48 and 49. Each lane contains 90 ng HK853 (A) Increasing concentrations of 48 competed with B-ATPγS only at concentrations greater than 800 µm (B) Increasing concentrations of 49 competed steadily with B- ATPγS. S25

26 Fig. S7 Rescue of HK853 activity in the presence of inhibitory concentrations of compounds 3, 48 and 49. Each lane contains 97 ng HK853. (A) A constant concentration of compound 48 (800 µm) does not show competitive effects on B-ATPγS activity-based labeling of HK853. The defects in lanes 2-4 are due to a deformity in the gel (B) A constant concentration of compound 49 (800 µm) prevents approximately 60% B-ATPγS labeling of HK853. The difference between lanes 1 and 8 illustrates the effects of 49 on the ability of HK853 to autothiophosphorylate. (C) NH125 (3) is marketed as an HK inhibitor, and inhibition of HK autophosphorylation was shown in previous studies 19. A constant concentration of NH125 (3) (800 µm) prevents approximately 60% B-ATPγS labeling of HK853. The difference between lanes 1 and 8 illustrates the effects of NH125 (3) on the ability of HK853 to autothiophosphorylate. S26

27 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications Fig. S8 Aggregation analysis of HK853 in the presence of compounds 3, 48 and 49. (A) Native-PAGE shows that HK853 aggregates in the presence of greater than 100 µm of compound ng of HK853 was loaded in each lane. (B) Crosslinking experiments also show 48 causes aggregation at concentrations greater than 100 µm. 85 ng HK853 was loaded per lane. The addition of Triton X-100 prevents 48induced aggregation (C) Native-PAGE shows that HK853 aggregates in the presence ~50 µm ng of HK853 was loaded in each lane. (D) Crosslinking experiments also show that 49 causes aggregation at concentrations of approximately 50 µm. 85 ng HK853 was loaded per lane. The addition of Triton X-100 prevents 49-induced aggregation. (E) Native-PAGE shows that HK853 aggregates in the presence of 50S27

28 100 µm NH125 (3). 95 ng of HK853 was loaded in each lane. The addition of Triton X-100 prevents NH125 (3)-induced aggregation (F) Crosslinking experiments also show that NH125 (3) causes aggregation at concentrations greater than 10 µm. 85 ng HK853 was loaded per lane. The addition of Triton X-100 prevents NH125 (3)-induced aggregation. Fig. S9 Triton X-100 restoration of HK853 activity labeling. Since NH125 (3) was proposed to be a nonspecific colloidal aggregator 20, Triton X-100 was added to prevent NH 125-induced aggregation. As judged by coomassie staining, there is even loading of protein in lanes 3 and 4. Lane 3 in the fluorescence gel shows a drastic decrease in activity-based labeling; however, restoration of labeling was demonstrated when detergent was added in lane 4. This supports that the mechanism of HK inhibition by NH125 (3) is through aggregation. Additionally, Triton X-100 was used in aggregation experiments to restore HK853 to native oligomeric states. 90 ng HK853 was loaded per lane. S28

29 Fig. S10 Competition and rescue of HK853 activity in the presence of compound 50 (A) Increasing concentrations of fragment 50 did not show competition with B-ATPγS. Each lane contains 90 ng HK853. The irregular band shapes in lanes 2 and 3 are due to a deformity in the gel. (B) A constant concentration of fragment 50 (800 µm) has no effect on B-ATPγS activity-based labeling of HK ng of HK853 was loaded in each lane. S29

30 Fig. S11. Analysis of guanidine for HK853 aggregation and competition with activity-based probe. Since guanidinium salts can be used to denature proteins, we wanted to be sure that the guanidine group was not causing denaturation, thus decreasing the activity of HK853. The molar ratio of guanidine:guanidine-hcl is ~0.6:1. Guanidine-HCl was used to prepare known concentrations of guanidine, and the final ph was 8.5. (A) Native-PAGE shows that guanidine does not cause aggregation at concentrations as high as 2 mm. 85 ng HK853 was loaded per lane. (B) Increasing concentrations of guanidine were not shown to affect the autothiophosphorylation of HK853, ensuring that decreases in labeling in activity-based experiments were not due to guanidine-induced denaturation. Each lane contains 90 ng HK853. The irregular band shapes in lanes 2-4 are due to a deformity in the gel. S30

31 Fig. S12. Aggregation analysis of HK853 in the presence of the fragment 50. (A) Native-PAGE shows that HK853 does not aggregate in the presence of the fragment ng of HK853 was loaded in each lane. (B) Crosslinking experiments also show that the fragment 50 does not cause aggregation. 85 ng HK853 was loaded per lane. S31

32 Fig. S13 Competition of HK853 activity in the presence of guanidine fragments 50 and 51. Each lane contains 90 ng HK853 (A) Increasing concentrations of 50 competed with B-ATPγS (B) Increasing concentrations of 51 competed with B-ATPγS. Table S2. Competitive ABPP using 2 µm B-ATPγS (probe) and increasing concentrations of 51 (competitor) with HK853. Bands correlate to Fig. S13 B. 51 (mm) Integrated Density of Band Fluorescence (AU) S32

33 Figure S14. Aggregation analysis of HK853 in the presence of the modified fragment 51. (A) Native- PAGE shows that HK853 aggregates slightly at concentrations between 4 and 8 mm of ng of HK853 was loaded in each lane. (B) Crosslinking experiments also show that compound 51 causes subtle aggregation at concentrations in the low micromolar range only. 85 ng HK853 was loaded per lane. S33

34 Docking-Based Virtual Screening General All molecular modeling operations were performed using SYBYL X2.0 on a quad-core Intel Core i3 workstation operating at 3.06 GHz equipped with 4 GB 1333MHz DDR 3 RAM. Visualization of docked poses was accomplished using the latest available release of UCSF Chimera. Docking of Compounds. The active sites of all receptors were defined based on the coordinates of the co-crystallized ligand and the Protomol Generation Mode in Surflex using default settings. A portion of the co-crystallized ligand was retained and used as a guide for the placement of compounds into the active site during docking. The Surflex-Dock parameters were configured as follows: Max Conformations per Fragment: 10 Max Number of Rotatable Bonds per molecule: 50 Activated: Consider Ring Flexibility Density of Search: 6.00 Maximum Number of Poses per Ligand: 10 Minimum RMSD Between Final Poses: 0.5 All other parameter were kept at their default values S34

35 In Vitro Activity Assays General Methods and Information Reagents were obtained from J.T. Baker, Mallinkrodt, Sigma, IBI, VWR, EMD Biosciences, Bio-Rad and Fisher. BODIPY-FL-ATPγS was purchased from Invitrogen, NH125 from Tocris Bioscience, and BS 3 -d 0 from Thermo Scientific. Experimental Methods HK853 Construct (Thermotoga maritima) The HK853 histidine kinase protein construct was generated as described previously 21 PCR Site-Directed Mutagenesis for Generation of HK853 D411A Construct 22 The DNA synthesized for wild-type HK853 was used as a template. Sense and antisense primers that originally coded for D411 were altered to alanine (See Table S3). Primers were ordered from New England Biolabs. Two reactions were prepared in PCR tubes: 2.5 ng HK853 wild-type DNA template, 2.5 µl 2.5 mm dntps, 2.5 µl 10 X Pfu buffer, µl nuclease-free water, and 0.25 µl 100 X Pfu. To one tube, 2.5 µl of 5 µm mutant sense primer (FHJ088) and 2.5 µl of 5 µm outermost wild-type antisense primer were added. To the other, 2.5 µl of 5 µm outermost wild-type sense primer and 2.5 µl of mutant antisense primer (FHJ089) were added. The final reaction volumes were 25 µl. The PCR reaction was 95 C for 60 s; 30 cycles of 95 C for 30 s, 56 C for 120 s, and 72 C for 90 s; and 72 C for 360 s. To amplify the mutated template, 0.5 µl of product from both the first and second tubes were mixed with 5.0 µl 2.5 mm dntps, 5.0 µl of 5 µm outermost sense primer, 5.0 µl of 5 µm outermost antisense primer, 5.0 µl of 10 X Pfu buffer, 28.5 µl nuclease-free water, and 0.5 µl 100 X Pfu to a total volume of 50 µl. The same PCR method was run. PCR product was purified, digested, and ligated into the p-his-parallel vector as before. 23 The DNA sequence was confirmed as successful through sequencing at the Indiana Molecular Biology Institute. Additionally, transformation of p-his-parallel-hk853 D411A S35

36 into E. coli strain BL21 (DE3)Rosetta, plyss, and subsequent protein overexpression and purification were performed as described previously 21. Overexpressed mutant protein is shown in Fig. S15, and the final protein properties are shown in Table S4. Table S3. Mutant primers used in PCR site-directed mutagenesis to generate HK853 D411A Primer Sequence Comments KEW026 TGAGAAAGACGGTGGTGTGCTGATCATCGTGGAGGATAATG Wild-type, sense, Asp KEW027 GGTCCGGGATGCCGATACCATTATCCTCCACGATGATCAG Wild-type, antisense, Asp FHJ088 TGAGAAAGACGGTGGTGTGCTGATCATCGTGGAGGCGAATG Mutant, sense, Ala FHJ089 GGTCCGGGATGCCGATACCATTCGCCTCCACGATGATCAG Mutant, antisense, Ala Figure S15. HK853 D411A overexpression. The protein ladder is on the left, non-induced E. coli lysate is in lanes 1 and 2 (two separate cultures), and induced E. coli lysate is in lanes 3 and 4 (two separate induced cultures). S36

37 Table S4. Final protein properties of the HK853 D411A mutant. Gray residues represent a polyhistidine tag coded by the p-his-parallel vector. The red residue is the mutated alanine. Values for pi and extinction coefficient are estimated. Circular Dichroism (CD) Spectroscopy of HK853 Proteins 24, 25 Previous CD methods were used as guidelines for this procedure. Using a Jasco J-715 CD spectropolarimeter, CD spectra were acquired for purified HK853 wild-type and D411A proteins. Proteins were exchanged into 10 mm potassium phosphate, ph 7.5, four times using 0.5-mL 10K Amicon Ultra centrifugal filters (Millipore). The Bio-Rad DC Protein Assay was used to determine protein concentrations, which were mg/ml for HK853 wild-type and mg/ml for HK853 D411A. Buffer and protein solutions were filtered with 0.22-µm Ultrafree-MC centrifugal filters (Millipore) to ensure the removal of any particulates that could interfere with CD readings. Protein solutions were loaded into a 0.1-cm quartz cuvette (Hellma), and spectra were obtained at 25 C. Each spectrum was measured in triplicate with the following parameters: standard (100 mdeg) sensitivity, nm range, 0.5 nm data pitch, continuous scanning mode, scanning speed of 100 nm/min, response of 1 s, 1.0-nm bandwidth, and an accumulation of 4 scans. Spectra were smoothed using a Savitsky-Golay filter (15- point smoothing window). Averaged buffer spectra were subtracted from the protein spectra. The CD data in millidegrees were used to calculate mean residue ellipticity, [θ], according to the following equation: [θ]= (millidegrees)/(path length in mm x concentration in M x number of amino acid residues). 25 The final units for mean residue ellipticity were deg cm -1 dmol -1. Additionally, data from each CD spectrum (in millidegrees) were submitted to Dichroweb for secondary structure analysis using SELCON3 and reference set Values for helices, strands, and turns from each spectrum were averaged, and error S37