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Table III.

Summary data for various mutants including amino acid properties, as stated for each mutation

Variant	Volume	Hydrophobicity		τ_act		τ_deact	V_1/2 of activation	Slope
	Å³		Charge	20 mV	at V_1/2	−50 mV
				ms	ms	ms	mV
D202	111.1	−3.5	Negative	729	729	126	17.7 ± 2.4	17.2 ± 1.0
D202N	114.1	−3.5	Neutral	400	392	11	41.5 ± 2.9^a	14.0 ± 1.5
D202H	153.2	−3.2	Pos/Neu	607	600	33	34.3 ± 5.3^b	14.5 ± 2.0
D202A	88.6	1.8	Neutral	644	604	44	29.8 ± 2.9	20.0 ± 2.2
D202E	138.4	−3.5	Negative	728	739	42	72.3 ± 3.4^a	17.2 ± 1.9
D202M	162.9	1.9	Neutral	738	799	28	41.2 ± 2.9^a	14.6 ± 1.5
D202K	168.6	−3.9	Positive	878	878	61	39.5 ± 3.9^a	11.5 ± 1.4^b
I204	166.7	3.8		729	729	126	17.7 ± 2.4	17.2 ± 1.0
I204M	162.9	1.9		845	768	82	53.8 ± 2.6^a	16.2 ± 0.6
I204F	189.9	2.8		5,287^c	3,083^c	54	71.0 ± 3.9^a	18.5 ± 2.0
I204A	88.6	1.8		1,511	925	170	36.4 ± 3.3^a	12.0 ± 1.1
I204N	114.1	−3.5		1,802	1,375	88	50.6 ± 9.8^a	16.0 ± 1.9
V205	140	4.2		729	729	126	17.7 ± 2.4	17.2 ± 1.0
V205M	162.9	1.9		1,081	825	53	37.0 ± 5.4^a	17.3 ± 2.2
V205L	166.7	3.8		333	507	75	0 ± 3.3	9.7 ± 0.6^a
V205A	88.6	1.8		915	1,332	52	68.8 ± 7.7^a	12.3 ± 2.0
S209	89	−0.8		729	729	126	17.7 ± 2.4	17.2 ± 1.0
S209F	189.9	2.8		496	1,902	430	−31.0 ± 2.3^a	14.9 ± 1.2
S209M	162.9	1.9		617	1,666	420	−22.6 ± 3.7^a	15.0 ± 0.9
S209A	88.6	1.8		295	650	302	−16.3 ± 3.9^a	15.0 ± 1.2
S209L	166.7	3.8		803	2,357	352	−30.1 ± 4.7^a	15.7 ± 2.3
V215	140	4.2		729	729	126	17.7 ± 2.4	17.2 ± 1.0
V215M	162.9	1.9		448	340	55	37.9 ± 4.3^b	14.4 ± 0.6
V215A	88.6	1.8		501	564	331	8.4 ± 2.3	14.0 ± 1.0
V215L	166.7	3.8		459	411	85	27.9 ± 4.3	13.6 ± 1.5
V215P	112.7	−1.6		593	551	29	55.7 ± 4.4^a	12.2 ± 0.9

Variant	Volume	Hydrophobicity		τ_act		τ_deact	V_1/2 of activation	Slope
	Å³		Charge	20 mV	at V_1/2	−50 mV
				ms	ms	ms	mV
D202	111.1	−3.5	Negative	729	729	126	17.7 ± 2.4	17.2 ± 1.0
D202N	114.1	−3.5	Neutral	400	392	11	41.5 ± 2.9^a	14.0 ± 1.5
D202H	153.2	−3.2	Pos/Neu	607	600	33	34.3 ± 5.3^b	14.5 ± 2.0
D202A	88.6	1.8	Neutral	644	604	44	29.8 ± 2.9	20.0 ± 2.2
D202E	138.4	−3.5	Negative	728	739	42	72.3 ± 3.4^a	17.2 ± 1.9
D202M	162.9	1.9	Neutral	738	799	28	41.2 ± 2.9^a	14.6 ± 1.5
D202K	168.6	−3.9	Positive	878	878	61	39.5 ± 3.9^a	11.5 ± 1.4^b
I204	166.7	3.8		729	729	126	17.7 ± 2.4	17.2 ± 1.0
I204M	162.9	1.9		845	768	82	53.8 ± 2.6^a	16.2 ± 0.6
I204F	189.9	2.8		5,287^c	3,083^c	54	71.0 ± 3.9^a	18.5 ± 2.0
I204A	88.6	1.8		1,511	925	170	36.4 ± 3.3^a	12.0 ± 1.1
I204N	114.1	−3.5		1,802	1,375	88	50.6 ± 9.8^a	16.0 ± 1.9
V205	140	4.2		729	729	126	17.7 ± 2.4	17.2 ± 1.0
V205M	162.9	1.9		1,081	825	53	37.0 ± 5.4^a	17.3 ± 2.2
V205L	166.7	3.8		333	507	75	0 ± 3.3	9.7 ± 0.6^a
V205A	88.6	1.8		915	1,332	52	68.8 ± 7.7^a	12.3 ± 2.0
S209	89	−0.8		729	729	126	17.7 ± 2.4	17.2 ± 1.0
S209F	189.9	2.8		496	1,902	430	−31.0 ± 2.3^a	14.9 ± 1.2
S209M	162.9	1.9		617	1,666	420	−22.6 ± 3.7^a	15.0 ± 0.9
S209A	88.6	1.8		295	650	302	−16.3 ± 3.9^a	15.0 ± 1.2
S209L	166.7	3.8		803	2,357	352	−30.1 ± 4.7^a	15.7 ± 2.3
V215	140	4.2		729	729	126	17.7 ± 2.4	17.2 ± 1.0
V215M	162.9	1.9		448	340	55	37.9 ± 4.3^b	14.4 ± 0.6
V215A	88.6	1.8		501	564	331	8.4 ± 2.3	14.0 ± 1.0
V215L	166.7	3.8		459	411	85	27.9 ± 4.3	13.6 ± 1.5
V215P	112.7	−1.6		593	551	29	55.7 ± 4.4^a	12.2 ± 0.9

Hydrophobicity was based on the Kyte Doolittle scale, and other values were obtained as indicated in the legend to Fig. 2.

Significant difference from WT (P < 0.01).

Significant difference from WT (P < 0.05).

τ_act for I204F activation was measured at 6 s rather than 2 s due to this process being very slow. Only V_1/2 values were tested for significance.