Fig. 2.

Hydroxylamine sensitivity and site-directed mutagenesis of the palmitoylated Kv1.1 channel protein. (A) Immunoprecipitated proteins were treated with buffer alone (-) or buffer containing 1 M hydroxylamine at pH 7 (+) before SDS/PAGE and autoradiography (Upper) or Western blotting analysis (Lower). (B) Comparison of palmitoylation of the Kv1.1 WT (WT-Kv1.1) with the C35A, C36A double mutant metabolically labeled with [³H]palmitate. Sf9 cells expressing either WT protein (WT-Kv1.1) or the C35A, C36A double mutant were metabolically labeled with [³H]palmitate, immunoprecipitated, and subjected to SDS/PAGE and autoradiography (Upper) or Western blotting (Lower). (C) Comparison of the palmitoylation of the Kv1.1 C243A mutant to palmitoylation of the C-terminal Kv1.1 deletion mutant D478-495 was performed after metabolic labeling with [³H]palmitate or [³⁵S]methionine, immunoprecipitation, and autoradiography.