Supporting Information for Studies Leading to the Development of a Highly Selective Colorimetric and Fluorescent Chemosensor for Lysine Ying Zhou, a Jiyeon Won, c Jin Yong Lee, c * and Juyoung Yoon a, b * a Department of Chemistry and Nano Science; b Department of Bioinspired Science, Ewha Womans University, Seoul 120-750, Korea; c Department of Chemistry, Sungkyunkwan University, Suwon 440-746, Korea jyoon@ewha.ac.kr;jinylee@skku.edu Experimental Section..... S2 page Figure S1, Figure S2.. S3 page Figure S3, Figure S4....S4 page Figure S5, Figure S6........S5 page Figure S7........S6 page Figure S8, Figure S9........S7 page Figure S10........S8 page Figure S11....S9 page Figure S12.........S10 page Figure S13.......S11 page Figure S14......S12 page Figure S15, Figure S16.......S13 page Figure S17, Figure S18...S14 page Figure S19, Figure S20...S15 page Figure S21, Figure S22...S16 page Figure S23, Figure S24...S17 page Figure S25, Figure S26...S18 page S1
Figure S27, Figure S28....S19page Figure S29, Figure S30........S20 page Figure S31, Figure S32........S21 page Figure S33, Figure S34.......S22 page Figure S35, Figure S36,.......S23 page Figure S37, Figure S38,.......S24 page Experimental Section General methods Unless otherwise noted, materials were obtained from commercial suppliers and were used without further purification. Flash chromatography was carried out on silica gel (230-400 mesh). 1 H NMR and 13 C NMR spectra were recorded using 250 MHz. Chemical shifts were expressed in ppm and coupling constants (J) in Hz. Mass spectra were obtained using JEL JMS-700 Mstation spectrometers. Preparation of amino acids and other testing target solutions Stock solutions (0.1 M) of the amino acids, such as lysine(lys), histidine (His), leucine (Leu), isoleucine (Ile), methionine (Met), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), valine (Val), asparagines (Asn), aspartic acid (Asp), arginine (Arg), alanine (Ala), serine (Ser), glutamine (Gln), glutamic acid (Glu), glycine (Gly), phenylalanine (Phe) and cysteine (Cys), and butylamine, ethylenediamine, 1, 3-diaminopropane, 4- aminobutyric acid, N-(tert-butoxycarbonyl) D lysine, peptide (Lys-Gly-Lys), 2- hydroxy-1-naphthaldehyde were prepared in DW. Stock solutions of host 1 were prepared in CH 3 CN/HEPES (0.01 M, ph = 7.4) (1: 9). For all measurements, excitation and emission slit widths were 5 and 10 nm, respectively. S2
1 H NH 2 NH 2 Figure S1. UV spectra of 1 (20 μm) in CH 3 CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition of 100 equiv. of D-lysine (0.1M in DW). 1 H NH 2 NH 2 Figure S2. Fluorescent spectra of 1 (20 μm) in CH3CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition 100 equiv. of L-lysine (0.1M in DW). Excitation wavelength was 380 nm. S3
N H H 2 N H Figure S3. HRFAB mass spectrum of compound 2. 1 H 2 N Figure S4. Fluorescent spectra of 1 (20 μm) in CH3CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition 10 equiv. of butylamine (0.1M in DW). Excitation wavelength was 390 nm. S4
Figure S5. 1 H NMR spectra of 2 in DMS. Figure S6. 13 C NMR spectra of 2 in DMS. S5
Figure S7. 1 H NMR spectra of 1 in DMS in the presence of a different amount of lysinein D2. S6
Hc N Hb Figure S8. 1 H NMR spectra of 1-BA in DMS. Ha Hb Adding of 1 equv. Butylamine Adding of 1 equv. Butylamine 20 mins later Figure S9. 1 H NMR spectra of 1 in DMS in the presence of butylamine. S7
1. in 100 % DMS H 5 H 3 H 17 N H H 13 H 6 H 12 H 8 H 10 H 9 2. in DMS : D2 = 1: 1 3. in DMS : D2 2: 1 4. the sample of 3 (12 hours later ) with more DMS 5. add NaH solid to sample 4 Figure S10. The NMR data of compound 1 with butylamine in different systems. S8
H 18 H 18 H 17 H 19 H 19 H 21H21 H 22 N H 20 H 20 NH 2 H H 3 H H 5 H 13 H 6 H 12 H 8 H 9 H 10 Figure S11. 500 MHz HSQC spectrum of 1 in DMS. S9
Figure S12. 500 MHz CSY spectrum of 1 in DMS. S10
Figure S13. 500 MHz HMBC spectrum of 1 in DMS. S11
. Figure S14. 500 MHz NESY spectrum of 1 in DMS. S12
1 H NH 2 Figure S15. The NMR of compound 1 with excess leucine (Leu) in DMS-D 2 (2:1). 1 H 2 N H Figure S16. The NMR of compound 1 with excess alanine (Ala) in DMS-D 2 (2:1). S13
1 H 2 N NH 2 H (2:1). Figure S17. The NMR of compound 1 with excess glutamine (Gln) in DMS-D 2 1 H NH 2 H Figure S18. The NMR of compound 1 with excess glutamic acid (Glu) in DMS- D 2 (2: 1). S14
Figure S19. The UV spectra of 1 (20 μm) in CH 3 CN-HEPES buffer (0.01 M, p H 7.4) (1: 9, v/v) after addition of 50 equiv. of L-Lys (0.1 M in DW), 10 equi v. of butylamine. Inset: picture of a solution of 1 only (2), and after addition of 50 equiv. of L-Lys (1), 10 equiv. of butylamine (3). (a) Within 10 min after th e addition. Inset: The time dependent change in the absorbance of 1 (100 μm) u pon addition of 70 equiv of L-Lys. (b) After 10 h of the addition. Inset: The ti me dependent change in the absorbance of 1 (100 μm) at 486 nm upon addition of 70 equiv of butylamine. Figure S20. UV spectra of 1 (20 μm) in CH 3 CN-HEPES buffer (0.01 M, ph=7.4) (1:9, v/v) upon addition of 100 equiv. of N-(tert-butoxycarbonyl)-D-Lys. Inset: an absorbance versus time plot following addition of 70 equiv of N-(tert-butoxycarbonyl)- D-Lys to 1 (100 μm) in CH 3 CN-HEPES buffer (0.01 M, ph=7.4) (1:9, v/v). S15
1 H 2 N H Figure S21. UV spectra of 1 (20 μm) in CH 3 CN-HEPES buffer (0.02 M, ph=7.4) (1: 9, v/v) upon addition of 30 equiv. of 4-aminobutyric acid (0.1M in DW). 1 H 2 N H Figure S22. Fluorescent spectra of 1 (20 μm) in CH3CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition of 30 equiv. of 4-aminobutyric acid (0.1M in DW). Excitation wavelength was 390 nm. S16
H 1 HN NH 2 Figure S23. UV spectra of 1 (20 μm) in CH3CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition of 100 equiv. of N-(tert-butoxycarbonyl)-D-lysine (0.1M in DW). H 1 HN NH 2 Figure S24. Fluorescent spectra of 1 (20 μm) in CH 3 CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition of 100 equiv. of N-(tert-butoxycarbo nyl)-d-lysine (0.1M in DW). Excitation wavelength was 380 nm. S17
1 H 2 N N H H N H NH 2 NH 2 Figure S25. The UV spectra of 1 (10 μm) in CH 3 CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition of 60 equiv. of Lys-Gly-Lys (KGK) (0.1M in DW) 1 H 2 N N H H N H NH 2 NH 2 Figure S26. Fluorescent spectra of 1 (20 μm) in CH 3 CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition of 60 equiv. of Lys-Gly-Lys (KGK) (0.1M in DW). Excitation wavelength was 390 nm. S18
H H Figure S27. The UV spectra of 2-hydroxy-1-naphthaldehyde (10 μm) in CH 3 CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition of 50 equiv of L- Lysine (0.1M in DW). H H Figure S28. The UV spectra of 1, 8-Dihydroxyanthrone (10 μm) in CH 3 CN- HEPES buffer (0.01 M, ph=7.4) (2: 8, v/v) upon addition of 50 equiv of L- Lysine (0.1M in DW). S19
0.4 H 0.3 CH 3 Absorbance 0.2 0.1 0.0 300 400 500 600 700 800 Wavelength (nm) Figure S29. The UV spectra of 1-methoxypyrene-2-carbaldehyde (20 μm) in CH3CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition of 50 eq uiv of L-Lysine (0.1M in DW). 500 H Fluorescence Intensity 400 300 200 100 CH 3 0 350 400 450 500 550 600 Wavelength (nm) Figure S30. Fluorescent spectra of 1-methoxypyrene-2-carbaldehyde (20 μm) in CH3CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v) upon addition of 50 equiv. of Lys (0.1M in DW). Excitation wavelength was 330 nm. S20
110 100 T/ % 90 80 70 3437 3039 2862 2933 1180 1407 1411 844 N H H 2 N H 60 50 4000 3500 3000 2500 2000 1500 1000 500 σ/ cm -1 1631 1535 Figure S31. IR spectrum of compound 2. 110 100 T/ % 90 80 70 60 2933 1627 1013 1181 883 841 N H 50 40 4000 3500 3000 2500 2000 1500 1000 500 σ/ cm -1 Figure S32. IR spectrum of compound 1-BA. S21
0.5 0.4 Absorbance 0.3 0.2 0.1 498 nm 0.0 0 500 1000 1500 time/ sec Figure S33. The kinetics upon addition of 70 equiv of 4-aminobutyric acid to 1 (100 um) in CH 3 CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v). 0.7 0.6 498 nm 0.5 Absorbance 0.4 0.3 0.2 0.1 0.0 0 500 1000 1500 2000 2500 3000 time/ sec Figure S34. The kinetics upon addition of 70 equiv of N-(tert-butoxycarbonyl)- D-lysine to 1 (100 um) in CH 3 CN-HEPES buffer (0.01 M, ph=7.4) (1: 9, v/v). S22
scillation strength 1.2 1.0 0.8 0.6 0.4 0.2 1 2 2-W2 1-BA 0.0 200 300 400 500 600 700 wave length (nm) Figure S35. Calculated absorption spectra of 1, 2, 2-W 2, and 1-BA. Figure S36. Structures of 1, 2, 2-W 2 and 1-BA. S23
scillation strength 0.5 0.4 0.3 0.2 0.1 1-dimer 2-dimer 2-W2-dimer 1-BA-dimer P2-Lys-W2-dimer 0.0-0.1 200 300 400 500 600 700 wave length (nm) Figure S37. Calculated absorption spectra of the stacked dimers of 1, 2, 2-W 2, 1-BA, and P2-Lys-W 2. 1000 Fluorescence Intensity 800 600 400 200 0 CH 3 CN : HEPES = 1 : 4 CH 3 CN : HEPES = 1 : 7 CH 3 CN : HEPES = 1 : 9 CH 3 CN : HEPES = 1 : 2 CH 3 CN : HEPES = 1 : 1 CH 3 CN : HEPES = 3 : 1 100 % CH 3 CN 350 400 450 500 550 600 650 Wavelength (nm) Figure S38. The fluorescence spectra of compound 1 (20 μm) in different ratios CH 3 CN and HEPES buffer (0.01 M, ph 7.4). Excitation wavelength = 340 nm. S24