Table S2.
Summary of biHis sites in WT Protein G
| Location | Site | Mutation | ψ0* | ϕbiHis | ΔΔGeqMetal | ΔΔGeqbiHis | ΔΔGfMetal | m0 | m0Metal | βT† | βTMetal | Metal |
| WT | − | − | − | − | − | − | 1.69(2) | − | 0.83(1) | − | − | |
| β1 + β2 | a | N18H/K13H | −0.02(0) | 0.18(3) | 1.76(5) | −0.97(5) | −0.23(3) | 1.99(4) | 1.66(4) | 0.79(1) | 0.77(2) | Zn2+ |
| −0.10(2) | 1.30(7) | −1.11(7) | 1.77(4) | 0.78(2) | Ni2+ | |||||||
| b | I6H/E15H | −5.13(386) | 0.48(2) | 0.09(6) | −1.28(4) | −1.00(8) | 1.99(9) | 1.82(6) | 0.78(2) | 0.71(4) | Zn2+ | |
| c | K4H/E15H | −0.24(3) | 0.01(4) | 0.72(4) | 0.57(3) | −0.50(3) | 1.56(3) | 1.29(2) | 0.73(3) | 0.73(3) | Zn2+ | |
| d | K4H/T17H | −0.22(7) | ND | −1.43(3) | −0.21(3) | 0.11(3) | 1.63(2) | 1.57(3) | 0.72(3) | 0.71(3) | Zn2+ | |
| α | e | A24H/K28H | −0.34(8) | 0.18(5) | 0.38(4) | −0.73(5) | −0.22(3) | 1.71(6) | 1.45(4) | 0.81(2) | 0.81(2) | Zn2+ |
| −0.83(17) | 0.34(5) | −0.61(4) | 1.27(4) | 0.70(2) | Ni2+ | |||||||
| f | K28H/Q32H | 0.24(4) | 0.13(3) | 1.05(12) | −1.18(5) | 0.46(12) | 1.88(7) | 1.76(8)§ | 0.80(2) | 0.85(2) | Zn2+ | |
| −0.05(3) | 1.00(9) | −0.16(8) | 1.71(8)§ | 0.84(2) | Ni2+ | |||||||
| g | Q32H/D36H | 0.03(3) | 0.21(2) | 0.82(5) | −0.94(3) | 0.05(5) | 1.84(2) | 1.82(3)§ | 0.70(2) | 0.73(3) | Zn2+ | |
| β3 + β4 | h | D46H/T51H | 0.90(1)¶ | ND | ND | ND | ND | ND | ND | ND | ND | Zn2+ |
| i | T44H/T53H | −2.57(53) | 0.04(2) | −0.49(5) | −1.54(4) | 0.52(5) | 1.85(4) | 1.72(5)§ | 0.85(1) | 0.92(1) | Zn2+ | |
| 0.93(2) | −2.42(17) | −1.44(17) | 1.82(10)§ | 0.88(1) | Ni2+ | |||||||
| β1 + β4 | j | K4H/T51H | 0.17(2) | 0.70(1) | 1.30(10) | −2.34(5) | 0.51(9) | 1.59(7) | 1.70(6)§ | 0.74(3) | 0.75(3) | Zn2+ |
| k | I6H/T53H | 0.71(5) | 0.41(2) | 1.21(14) | −1.75(5) | 1.04(14) | 1.71(10) | 1.68(11)§ | 0.81(0) | 0.81(0) | Zn2+ | |
| l | N8H/T55H | 0.30(1) | 0.13(2) | 1.77(4) | −0.80(3) | 1.13(3) | 1.85(1) | 1.42(2) | 0.73(2) | 0.79(2) | Zn2+ | |
| β2 + α | m | T16H/Y33H | 0.24(2) | 0.33(2) | 1.47(5) | −1.67(6) | 0.77(3) | 1.95(3) | 1.45(4) | 0.73(3) | 0.71(3) | Zn2+ |
Values in parentheses represent SEs of the fitting and derived parameters, e.g., 0.18(3) = 0.18 ± 0.03. ND, not determined.
*
ψ0 value is calculated directly in the fit of the chevron data that includes data for a strongly folding and unfolding condition, xf and xu, respectively (Fig. S3).
†
βT = mf/m0, where mf and m0 are the denaturant dependences of ΔGf and ΔGeq, respectively (Materials and Methods).
§
mf is a shared parameter.
¶
Estimated from measuring the dependence of folding and unfolding kinetics in a Leffler style analysis (Fig. S2).