Figure 1.

Topology and in vitro lipidation of Cav3. (A) Cartoon depicting the predicted topology of monomeric Cav3 at the membrane, with the positions of the natively palmitoylated cysteine residues indicated (top). Cysteine thiols were first modified by the addition of excess 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB, Ellman’s reagent) (middle). 3-Thio-6-nitrobenzoate groups were then disulfide exchanged with thioalkyl chains in the presence of excess octanethiol (bottom). (B) Representative sodium dodecyl sulfate–polyacrylamide gel electrophoresis gel (Coomassie stain) of purified lipidated Cav3.