Table 4.

Rates of MTS-ET effects on TH6–TH7 segment mutant channels

Mutant cis pH mV kcis ktrans 
   M−1 s−1 M−1 s−1 
T300C 7.0 60 40 ± 13 (n = 4) NE (n = 2) 
T301C 6.2 60 15 ± 2 (n = 3) NE (n = 2) 
A302C 5.3 −20 8.1 ± 0.2 (n = 2) NE (n = 2) 
A303C 7.2 40–60 1,094 ± 91 (n = 3) NE (n = 3) 
L304C 6.6 60 68 ± 9 (n = 3) NE (n = 2) 
S305Ca 5.3 192 ± 40 79 ± 10 
I306C 5.3 −30 or 30b 4.4 ± 0.8 (n = 2) 5.6 ± 0.2 (n = 2) 
L307C 5.3 30–40 33 ± 9 (n = 3) 607 ± 377 (n = 6) 
G309C 5.3 20–60 40 ± 19 (n = 4) 750 ± 479 (n = 4) 
I310Cc 6.3 60 44 (n = 4) <2 (n = 3) 
S312Ca 5.3 7.4 ± 1 11.4 ± 2 
V313C 6.3 20–60 8 ± 4 (n = 4) NE (n = 2) 
M314C 6.3 20–60 10 ± 4 (n = 6) NE (n = 4) 
G315Cc 5.2 30–60 7 (n = 6) <0.5 (n = 4) 
I316C 5.2 40 32 ± 10 (n = 3) NE (n = 2) 
A317C 7.0 60 113 ± 72 (n = 3) NE (n = 2) 
Mutant cis pH mV kcis ktrans 
   M−1 s−1 M−1 s−1 
T300C 7.0 60 40 ± 13 (n = 4) NE (n = 2) 
T301C 6.2 60 15 ± 2 (n = 3) NE (n = 2) 
A302C 5.3 −20 8.1 ± 0.2 (n = 2) NE (n = 2) 
A303C 7.2 40–60 1,094 ± 91 (n = 3) NE (n = 3) 
L304C 6.6 60 68 ± 9 (n = 3) NE (n = 2) 
S305Ca 5.3 192 ± 40 79 ± 10 
I306C 5.3 −30 or 30b 4.4 ± 0.8 (n = 2) 5.6 ± 0.2 (n = 2) 
L307C 5.3 30–40 33 ± 9 (n = 3) 607 ± 377 (n = 6) 
G309C 5.3 20–60 40 ± 19 (n = 4) 750 ± 479 (n = 4) 
I310Cc 6.3 60 44 (n = 4) <2 (n = 3) 
S312Ca 5.3 7.4 ± 1 11.4 ± 2 
V313C 6.3 20–60 8 ± 4 (n = 4) NE (n = 2) 
M314C 6.3 20–60 10 ± 4 (n = 6) NE (n = 4) 
G315Cc 5.2 30–60 7 (n = 6) <0.5 (n = 4) 
I316C 5.2 40 32 ± 10 (n = 3) NE (n = 2) 
A317C 7.0 60 113 ± 72 (n = 3) NE (n = 2) 

The rates are given as mean ± SD, with n being the number of macroscopic experiments, except where indicated otherwise. The trans pH was 7.2 for all experiments. The His6-tag was removed from the T-domain for the experiments with A302C, S305C, I306C, and S312C. NE, no effect.

a

The rates for S305C and S312C are from the fitted lines in the graphs in Fig. 6. The error range was estimated by eye.

b

For I306C, kcis was measured at 30 mV and ktrans was measured at −30 mV.

c

A trans MTS-ET effect was observed in single-channel experiments with I310C and G315C, but it was too slow to be detected in macroscopic experiments. The rates given here were estimated from the single-channel waiting times. In some of the experiments with trans MTS-ET, the channel did not react after a long waiting time, so the value given for the rate is an upper limit. Here, n is the number of reaction events.

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