TABLE I

Analyzed Data of the Glycine Scanning Mutagenesis

Mutants
Activation
V1/2 (mV)
EC50 (μM)
G344A/S340G (−)  20.3 ± 8.0 (n = 6) 
G344A/I341G (+) −81.2 ± 7.3 (n = 7) 10.1 ± 1.5 (n = 6) 
G344A/G342G (G344A) (−)  24.6 ± 9.9 (n = 6) 
G344A/V343G (+) 6.4 ± 8.4 (n = 4) 48.4 ± 22.0 (n = 5) 
G344A/A344G (WT) (+) −54.2 ± 7.5 (n = 7) 13.5 ± 2.5 (n = 6) 
G344A/S345G (−)  47.5 ± 10.0 (n = 5) 
G344A/F346G (+) −105.6 ± 5.0 (n = 6) 13.3 ± 2.0 (n = 11) 
G344A/L347G (+) <−200 (n = 8) 24.1 ± 9.3 (n = 6) 
G344A/C348G (−)  51.1 ± 17.8 (n = 5) 
G344A/D349G Nonfunction Nonfunction Nonfunctional 
G344A/W350G (−)  28.2 ± 7.8 (n = 5) 
Mutants
Activation
V1/2 (mV)
EC50 (μM)
G344A/S340G (−)  20.3 ± 8.0 (n = 6) 
G344A/I341G (+) −81.2 ± 7.3 (n = 7) 10.1 ± 1.5 (n = 6) 
G344A/G342G (G344A) (−)  24.6 ± 9.9 (n = 6) 
G344A/V343G (+) 6.4 ± 8.4 (n = 4) 48.4 ± 22.0 (n = 5) 
G344A/A344G (WT) (+) −54.2 ± 7.5 (n = 7) 13.5 ± 2.5 (n = 6) 
G344A/S345G (−)  47.5 ± 10.0 (n = 5) 
G344A/F346G (+) −105.6 ± 5.0 (n = 6) 13.3 ± 2.0 (n = 11) 
G344A/L347G (+) <−200 (n = 8) 24.1 ± 9.3 (n = 6) 
G344A/C348G (−)  51.1 ± 17.8 (n = 5) 
G344A/D349G Nonfunction Nonfunction Nonfunctional 
G344A/W350G (−)  28.2 ± 7.8 (n = 5) 

Names of the analyzed mutants are shown in the first column. In the second column, (+) indicates recovery of the activation phase; (−) indicates no recovery. For the mutants with recovered activation phase, tail current analyses were performed, and the V1/2 values in the presence of 300 μM ATP (except 100 μM for G344A/V343G) are shown in the third column. V1/2 values for the G344A/L347G mutant were too hyperpolarized to analyze precisely. EC50 values for the [ATP] response relationship at −160 mV are indicated in the last column. G344A/D349G showed no detectable current.

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