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. Author manuscript; available in PMC: 2020 Aug 8.
Published in final edited form as: Cell. 2019 Jul 25;178(4):993–1003.e12. doi: 10.1016/j.cell.2019.06.031

Figure 2. Electrophysiological Recordings of NaVAb/WT, NaVAb/N49K, NaVAb/KAV and NaVAb/KAV Cysteine Mutants.

Figure 2

(A) Normalized conductance-voltage (G/V) relationships and Boltzmann fits for WT and each mutant. Sf9 cells expressing NaVAb KAV mutants (all C-terminal Δ28 truncation constructs) were stimulated with 50 ms pulses from 0–150 mV in 10 mV increments from a holding potential of −150 mV. Dashed: WT from (Gamal El-Din et al., 2013); Green, NaVAb/N49K, V1/2=−21.8±1.8 mV, k=7.9±1.0 mV, n=4; Gray: NaVAb/N49K/L109A/M116V (KAV) Δ28, V1/2=59.1±0.8 mV, k=7.7±0.8 mV, n=3; Blue: NaVAb/KAV/Q150C Δ28, V1/2=76.8±3.1 mV, k=10.0±0.9 mV, n=3; Red: NaVAb/KAV/G94C Δ28, V1/2=88.4±1.1 mV, k=10.6±0.4 mV, n=4; Purple: NaVAb/KAV/G94C/Q150C Δ28 in 50 mM DTT, V1/2=105.7±0.8 mV, k=10.2±0.2 mV, n=3; Black: NaVAb/KAV/G94C/Q150C Δ28 with no DTT, V1/2=n/a, k=n/a, n=5. Markers and error bars represent average G/Gmax ± standard error of the mean (SEM). Half activation (V1/2) and slope (k) values are averages of individual fits ± SEM. Curves = 1/(1+ê(( V1/2−Vm)/k)). Data points within 15 mV of Vrev were omitted to reduce noise.

(B) Representative current families of NaVAb/KAV Δ28 (gray), NaVAb/KAV/Q150C Δ28 (blue), NaVAb/KAV/Q94C Δ28 (red), and NaVAb/KAV/G94C/Q150C Δ28 in 50 mM DTT (purple). Transiently transfected Sf9 cells were held at −150 mV and stimulated for 50 ms to depolarized voltages in 10 mV increments. Scale bars represent 10 msec x 1 nAmp.

See also Figure S3.