Table 1. Stability, folding/unfolding rates, and Φ values of the mutants.

Mutation*	Whighq,† %	Nat. Cont.‡	Wlowq,§ %	τf,¶ ns	Nf∥	τu,** ns	Nu††	, kcal/mol‡‡	, kcal/mol‡‡	Φ‡‡§§
WT	21.4	19.3 ± 1.7	2.9	70 ± 10	92	67 ± 6	94
W2A	26.5	18.1 ± 2.3	3.5	107 ± 14	108	63 ± 6	114	–0.32 ± 0.15	–0.28 ± 0.13	0.87 ± 0.57
W2F	33.5	18.8 ± 2.2	3.4	106 ± 14	97	82 ± 8	103	–0.14 ± 0.16	–0.27 ± 0.13	—
W2L	24.9	18.2 ± 2.2	6.3	109 ± 16	101	63 ± 5	111	–0.34 ± 0.16	–0.30 ± 0.14	0.87 ± 0.57
W2V	23.6	18.3 ± 2.3	4.4	124 ± 17	95	62 ± 6	102	–0.43 ± 0.16	–0.38 ± 0.13	0.89 ± 0.45
W2Y	21.9	18.5 ± 2.4	6.4	129 ± 21	93	65 ± 6	98	–0.43 ± 0.16	–0.41 ± 0.14	0.95 ± 0.49
13A	19.9	18.7 ± 2.2	3.9	137 ± 18	92	64 ± 5	101	–0.48 ± 0.15	–0.44 ± 0.13	0.93 ± 0.40
13F	33.0	18.8 ± 2.1	3.3	121 ± 22	83	93 ± 8	91	–0.15 ± 0.17	–0.36 ± 0.15	—
13L	28.5	18.5 ± 2.4	3.9	119 ± 19	94	72 ± 7	101	–0.31 ± 0.17	–0.35 ± 0.14	1.1 ± 0.77
13M	30.2	18.9 ± 2.2	5.4	108 ± 19	94	81 ± 9	102	–0.16 ± 0.17	–0.29 ± 0.15	—
13V	37.2	18.6 ± 2.1	5.2	124 ± 18	75	109 ± 10	83	–0.06 ± 0.16	–0.38 ± 0.14	—
W10A	31.8	19.5 ± 2.1	5.0	161 ± 21	74	95 ± 10	79	–0.32 ± 0.16	–0.55 ± 0.13	1.7 ± 0.93
W10F	41.3	18.7 ± 2.2	3.8	77 ± 9	120	78 ± 6	127	0.04 ± 0.14	–0.06 ± 0.12	—
W10G	12.7	19.3 ± 2.2	3.1	212 ± 32	60	68 ± 9	69	–0.72 ± 0.17	–0.73 ± 0.14	1.0 ± 0.31
W10I	30.8	18.3 ± 2.1	6.0	129 ± 17	77	88 ± 9	83	–0.23 ± 0.16	–0.40 ± 0.13	—
W10L	20.8	18.8 ± 2.2	4.2	166 ± 22	81	58 ± 5	87	–0.67 ± 0.16	–0.57 ± 0.13	0.86 ± 0.28
W10M	18.4	19.0 ± 2.2	6.6	155 ± 21	82	52 ± 5	91	–0.68 ± 0.16	–0.52 ± 0.13	0.76 ± 0.26
W10V	17.2	17.8 ± 2.5	6.7	259 ± 40	57	65 ± 11	64	–0.88 ± 0.19	–0.86 ± 0.14	0.98 ± 0.26
W10Y	26.2	19.0 ± 2.1	3.5	118 ± 15	94	77 ± 7	98	–0.26 ± 0.15	–0.35 ± 0.13	—
Y11A	5.7	18.1 ± 2.0	2.3	249 ± 38	64	30 ± 3	71	–1.37 ± 0.17	–0.84 ± 0.14	0.61 ± 0.13
Y11F	33.1	19.1 ± 2.2	4.4	138 ± 20	73	112 ± 12	79	–0.11 ± 0.16	–0.45 ± 0.14	—
Y11L	14.8	18.6 ± 2.1	4.8	169 ± 23	76	54 ± 6	83	–0.72 ± 0.16	–0.58 ± 0.13	0.81 ± 0.26
Y11M	11.3	18.0 ± 2.2	3.5	152 ± 24	95	35 ± 3	105	–0.94 ± 0.16	–0.51 ± 0.14	0.54 ± 0.18
Y11V	5.7	17.0 ± 2.7	7.4
I18A	12.3	18.5 ± 2.3	2.4	168 ± 22	80	53 ± 6	88	–0.73 ± 0.16	–0.58 ± 0.13	0.79 ± 0.25
I18F	21.3	19.0 ± 2.0	3.2	159 ± 23	74	72 ± 8	83	–0.50 ± 0.17	–0.54 ± 0.14	1.1 ± 0.46
I18L	22.2	19.0 ± 2.2	4.4	145 ± 19	73	94 ± 9	81	–0.26 ± 0.16	–0.48 ± 0.13	—
I18M	28.9	18.8 ± 2.2	4.8	97 ± 15	99	77 ± 6	106	–0.13 ± 0.16	–0.22 ± 0.14	—
I18V	29.6	18.8 ± 2.3	3.2	124 ± 20	87	86 ± 9	93	–0.22 ± 0.17	–0.38 ± 0.14	—
Y19A	20.7	18.6 ± 2.4	7.4	123 ± 18	90	84 ± 8	95	–0.23 ± 0.16	–0.37 ± 0.14	—
Y19F	29.2	18.4 ± 2.2	3.8	130 ± 18	92	71 ± 7	98	–0.37 ± 0.16	–0.41 ± 0.13	1.1 ± 0.59
Y19L	30.0	18.3 ± 2.2	3.2	117 ± 17	83	88 ± 8	89	–0.17 ± 0.16	–0.34 ± 0.13	—
Y19M	17.5	18.5 ± 2.3	6.2	155 ± 26	68	97 ± 10	76	–0.28 ± 0.17	–0.52 ± 0.15	—

^*Mutants in italics are radical but acceptable and mutations in Roman are conservative (see Methods and ref. 24)

^†Statistical weight of the three most populated clusters with Q ≥ 16/24

^‡Average number of contacts in the three most populated clusters with Q ≥ 16/24

^§Statistical weight of the three most populated clusters with Q < 16/24

^¶Average folding time

^∥Number of folding events

^**Average unfolding time

^††Number of unfolding events

^‡‡The SD have been obtained by propagation of the error on τ_f and τ_u

^§§Dashes indicate unreliable Φ values because of kcal/mol. The reliable Φ values and the corresponding large stability changes (24) are bold. The multipoint Φ values are 0.77, 0.60, 0.79, 0.46, 0.72, and 1.23 for W2, I3, W10, Y11, I18, and Y19, respectively