Calcium-dependent membrane penetration is a hallmark of the C2 domain of cytosolic phospholipase A2 whereas the C2A domain of synaptotagmin binds membranes electrostatically.

C2 domains have been identified in a wide range of intracellular proteins, including lipid modifying enzymes, protein kinases, GTPases, and proteins involved in membrane trafficking. Many C2 domains bind membranes in a calcium-dependent manner. The first C2 domain from synaptotagmin I (SytIC2A) and the C2 domain from cytosolic phospholipase A2 (cPLA2C2) are among the best characterized C2 domains in terms of their structures and calcium binding. Here we demonstrate that the protein-lipid interaction is dramatically different for these two domains. Photolabeling with 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine ([125I]TID) in the presence of phospholipid vesicles indicates that cPLA2C2 penetrates into the hydrophobic region of the membrane. Hydrophobic surfaces on cPLA2C2 are exposed even in the absence of calcium, but only in its presence does the domain penetrate into the nonpolar core of the membrane. The interaction of SytIC2A with phospholipid membranes is primarily electrostatic with binding being abolished in 500 mM NaCl. Because soluble phospholipid head group analogues do not compete with binding of either SytIC2A or cPLA2C2 to vesicles, it is likely that membrane binding by these domains involves multiple interactions.