Table 2.

Kinetic parameters for 10-glycine insertion and disulfide crosslink mutants

Mutant	ln(kf)1M	ln(ku)5.5M	mf	mu	ΔGU–F	ΔΔGU–F
WT	2.36	2.44	1.02	0.54	3.95	—
10gly	0.997	2.30	1.20	0.44	2.86	−0.72
SS_red	2.75	3.38	1.05	0.33	2.50	−0.32
SS_ox	6.50	3.37	0.750	0.59	5.83	1.88
RT_red	2.65	2.37	1.18	0.39	3.47	0.21
RT_ox	3.81	0.62	0.78	0.42	4.98	1.92
NC_red	2.58	1.59	1.03	0.50	4.36	0.63
NC_ox	4.73	−1.30	0.766	0.80	8.70	3.58

Kinetics of folding and unfolding were followed by changes in tryptophan fluorescence on a stopped flow instrument at 295 K; k_f is reported in 1 M Gnd, and k_u is in 5.5 M Gnd to avoid extrapolation; m_f and m_u are the dependences of the folding and the unfolding rates, respectively, on Gnd. ΔG_U–F (free energy of unfolding) and ΔΔG_U–F (the difference in ΔG_U–F between the particular mutant protein and WT) were calculated from the kinetic parameters as described in the Methods section. Typical errors for the kinetic measurements are 1–10% as reported in ref. 13. Abbreviations are defined in the legend to Fig. 3.