Structural effects of LacI variants

Transcription

1 Structural effects of aci variants 1 of 6 10/9/2012 1:05 PM Structural effects of aci variants TMD Change[1] Y282D Reversion[2] Variant ssembly Y7 W/Wless[3] M42 I 62 W/Wless[3] I64 1 V H74 2 R /Wless[3] Protein Stability Chemical Modification Solvent Exposure Compared to WT S77 2 [4] NS more apolar KI: ess quenching/solvent exposure with IPTG change Q78 1 V V[5] C107 IPTG I83 1 K84 1,2 E R D S93 1 V94 1 V95 V96 1 S97 1 [6] High[7] Very high sensitivity lost Monoclonal Exposure WT6 WT Decreased Decreased UV Difference Bigger >300 nm Very small diff ll bigger[8] peaks ll bigger peaks

2 Structural effects of aci variants 2 of 6 10/9/2012 1:05 PM M98 1 I E100 1 W/Wless[3] N113 1 Q117 1 W/Wless[3] R118 Y T V F[9] D149 1 N V150 I S151 P[8] F161 3 S162 3 H163 3 E164 3 S191 F S193 1 R197 1 G[10] K W201[11] W22011 F226 W/Wless[3] M223 I T M232 I N246 3 S Q248 3 R 251 [12] Y273 W/Wless[3] D274 3 N[13] G K[14] Easily proteolyzed More exposed

3 tructural effects of aci variants 3 of 6 10/9/2012 1:05 PM T276 2 I D278 [15] C281[16] S F M I S/282 Y282[17] D Monomer F P284 S K290 2 Q291 2 D292 2 F293 2 W/Wless[3] M V P320 3 [8] V321 3 I S322 3 K325 W/Wless[3] R326[18] K E W S354 F References Bandyopadhyay, P. K. and C.-W. Wu (1979). "Heterogeneity of the Two Tryptophanyl Residues on the lac Repressor of Escherichia coli." rch.biochem.biophys. 195, No. 2:

4 tructural effects of aci variants 4 of 6 10/9/2012 1:05 PM Barry, J. K. and K. S. Matthews (1997). "igand-induced conformational changes in lactose repressor: a fluorescence study of single tryptophan mutants." Biochemistry 36(50): Burns,. E.,. H. Maki, et al. (1992). "Characterization of the two tryptophan residues of the lactose repressor from Escherichia coliby phosphorescence and optical detection of magnetic resonance." Biochemistry 32: Chakerian,. E. and K. S. Matthews (1991). "Characterization of mutations in oligomerization domain of ac repressor protein." J Biol Chem 266(33): Chakerian,. E., M. Pfahl, et al. (1985). " mutant lactose repressor with altered inducer and operator binding parameters." J Mol Biol 183(1): Chang, W.-I., P. Barrera, et al. (1994). "Identification and characterization of aspartate residues that play key roles in the allosteric regulation of a transcription factor: spartate 274 is essential for inducer binding in lac repressor." Biochemistry 33: Chang, W. I. and K. S. Matthews (1995). "Role of sp274 in lac repressor: Diminished sugar binding and altered conformational effects in mutants." Biochemistry 34(28): Chang, W. I., J. S. Olson, et al. (1993). "ysine 84 is at the subunit interface of lac repressor protein." J Biol Chem 268(23): Chen, J. and K. S. Matthews (1992). "T41 mutation in lac repressor is Tyr sp." Gene 111(1): Chou, W. Y. and K. S. Matthews (1989). "Mutation in hinge region of lactose repressor protein alters physical and functional properties." J Biol Chem 264(11): Daly, T. J. and K. S. Matthews (1986). "Characterization and modification of a monomeric mutant of the lactose repressor protein." Biochemistry 25(19): Dong, F., S. Spott, et al. (1999). "Dimerisation mutants of ac repressor. I. monomeric mutant, 251, that binds ac operator DN as a dimer." J Mol Biol 290(3): Flynn, T. C.,. Swint-Kruse, et al. (2003). "llosteric transition pathways in the lactose repressor protein core domains: asymmetric motions in a homodimer." Protein Sci 12(11): Gardner, J.. and K. S. Matthews (1990). "Characterization of two mutant lactose repressor proteins containing single tryptophans." J Biol Chem 265(34): i,. and K. S. Matthews (1995). "Characterization of Mutants ffecting the KRK Sequence in the Carboxyl-terminal Domain of lac Repressor." J.Biol.Chem. 270,no.18: Nichols, J. C. and K. S. Matthews (1997). "Combinatorial mutations of lac repressor. Stability of monomer-monomer interface is increased by apolar substitution at position 84." J Biol Chem 272(30): Ozarowski,., J. K. Barry, et al. (1999). "igand-induced conformational changes in lactose repressor: a phosphorescence and ODMR study of single-tryptophan mutants." Biochemistry 38(21): Royer, C.., J.. Gardner, et al. (1990). "Resolution of the fluorescence decay of the two tryptophan residues of lac repressor using single tryptophan mutants." Biophys J 58:

5 Structural effects of aci variants 5 of 6 10/9/2012 1:05 PM Schmitz,., U. Schmeissner, et al. (1976). "Mutations affecting the quaternary structure of the lac repressor." J Biol Chem 251(11): Spott, S., F. Dong, et al. (2000). "Dimerisation mutants of ac repressor. II. single amino acid substitution, D278, changes the specificity of dimerisation." J Mol Biol 296(2): Spotts, R. O.,. E. Chakerian, et al. (1991). "rginine 197 of lac repressor contributes significant energy to inducer binding. Confirmation of homology to periplasmic sugar binding proteins." J Biol Chem 266(34): Swint-Kruse,., C. R. Elam, et al. (2001). "Plasticity of quaternary structure: twenty-two ways to form a aci dimer." Protein Sci 10(2): Swint-Kruse,., H. Zhan, et al. (2003). "Perturbation from a distance: mutations that alter aci function through long-range effects." Biochemistry 42(47): Swint-Kruse,., H. Zhan, et al. (2005). "Integrated insights from simulation, experiment, and mutational analysis yield new details of aci function." Biochemistry 44(33): Footnotes [1] Number indicates whether residue participates in pathways 1, 2, or 3. (Flynn, Swint-Kruse et al. 2003) [2] (Swint-Kruse, Elam et al. 2001) [3] (Barry and Matthews 1997) [4] ll data: (Chou and Matthews 1989) [5] ll data: (Chakerian, Pfahl et al. 1985) [6], E, R, : (Chang, Olson et al. 1993) [7], : (Nichols and Matthews 1997) [8], : (Swint-Kruse, Zhan et al. 2005) [9] (Swint-Kruse, Zhan et al. 2003) [10] (Spotts, Chakerian et al. 1991) [11] (Bandyopadhyay and Wu 1979; Gardner and Matthews 1990; Royer, Gardner et al. 1990; Burns, Maki et al. 1992; Barry and Matthews 1997; Ozarowski, Barry et al. 1999) [12] (Dong, Spott et al. 1999)

6 Structural effects of aci variants 6 of 6 10/9/2012 1:05 PM [13] (Chang and Matthews 1995) [14] (Chang, Barrera et al. 1994) [15] OTS OF OTHER MUTTIONS IN THIS PPER (Spott, Dong et al. 2000) [16] (Chakerian and Matthews 1991) [17] (Schmitz, Schmeissner et al. 1976; Daly and Matthews 1986; Chakerian and Matthews 1991; Chen and Matthews 1992) [18] (i and Matthews 1995)