Regulation of recombinant CaV3.1 protein membrane expression by Cdk5.
2015-03-11T03:33:08Z (GMT) by
<p>A) Representative macroscopic current traces recorded from HEK-293 cells transiently transfected with a plasmid encoding Ca<sub>V</sub>3.1 channels alone or in conjunction with plasmids encoding Cdk5 and p35. Currents were elicited by depolarizing steps from a <i>V</i><sub>h</sub> of −80|mV to-30|mV. Ba<sup>2+</sup> (2 mM) was used as the charge carrier. B) Conductance-voltage (<i>G-V</i>) curves were calculated for each cell. An increase in conductance was observed in Cdk5/p35-coexpressing cells. C) Cdk5/p35 overexpression in HEK-293 cells stably expressing Ca<sub>V</sub>3.1 channels did not affect the voltage dependence of current activation and steady-state inactivation. D) Comparison of the time constant of current activation (τ<sub>act</sub>) and inactivation (τ<sub>inact</sub>) at different membrane voltages in untransfected (control) and Cdk5/p35-coexpressing cells. E) Representative cell surface protein extraction assay followed by Western blot using an anti-Ca<sub>V</sub>3.1 specific antibody. F) Densitometric quantification of three repetitions of the experiment shown in E. Asterisk denotes significant differences (P>0.05) between cells expressing the Ca<sub>V</sub>3.1 channels only, and cells transfected with the channels plus the Cdk5/p35 complex.</p>