AI Article Synopsis
- C2 domains (C2s) are specialized protein modules in eukaryotic cells that interact with cell membranes.
- C2s can be divided into Ca(2+)-dependent and Ca(2+)-independent types, each engaging with different lipid molecules through various binding methods.
- The protocol outlines a biochemical method to produce and purify these C2 domains attached to GST to study their interactions with lipid partners.
Article Abstract
C2 domains (C2s) are regulatory protein modules identified in eukaryotic proteins targeted to cell membranes. C2s were initially characterized as independently folded Ca(2+)-dependent phospholipids binding domains; however, later studies have shown that C2s have evolutionarily diverged into Ca(2+)-dependent and Ca(2+)-independent forms. These forms interact and regulate their affinity to diverse lipid species using different binding mechanisms. In this protocol we describe a biochemical approach to produce, purify, and solubilize functional C2 domains bound to GST for the identification of their putative Ca(2+)-dependent and Ca(2+)-independent lipid-binding partners.
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