Selective fluorophore-assisted light inactivation of voltage-gated calcium channels

초록

Fluorophore-assisted light inactivation (FALI) is an investigative tool to inactivate fluorescently labeled proteins by a mechanism of in situ photodestruction. We found that Ca(v)1.2 (L-type) and Ca(v)3.1 (T-type) calcium channels, labeled by genetic fusion with GFP derivatives, show differential sensitivity to FALI. Specifically, FALI silences Ca(v)1.2 calcium channels containing EYFP-labeled alpha(1C) subunits but does not affect the EYFP-alpha(1G) Ca(v)3.1 calcium channels or Ca(v)1.2 channels containing EYFP-labeled beta subunits. Our findings limit the applicability of acceptor photobleaching for the measurements of FRET but open an opportunity to combine the fluroscent imaging of the live cell expressing labeled calcium channels with selective functional inactivation of their specifics subsets.

키워드