title: C2 domain membrane penetration by group IVA cytosolic phospholipase A₂ induces membrane curvature changes.
abstract: Group IVA cytosolic phospholipase A(2) (cPLA(2)α) is an 85 kDa enzyme that regulates the release of arachidonic acid (AA) from the sn-2 position of membrane phospholipids. It is well established that cPLA(2)α binds zwitterionic lipids such as phosphatidylcholine in a Ca(2+)-dependent manner through its N-terminal C2 domain, which regulates its translocation to cellular membranes. In addition to its role in AA synthesis, it has been shown that cPLA(2)α promotes tubulation and vesiculation of the Golgi and regulates trafficking of endosomes. Additionally, the isolated C2 domain of cPLA(2)α is able to reconstitute Fc receptor-mediated phagocytosis, suggesting that C2 domain membrane binding is sufficient for phagosome formation. These reported activities of cPLA(2)α and its C2 domain require changes in membrane structure, but the ability of the C2 domain to promote changes in membrane shape has not been reported. Here we demonstrate that the C2 domain of cPLA(2)α is able to induce membrane curvature changes to lipid vesicles, giant unilamellar vesicles, and membrane sheets. Biophysical assays combined with mutagenesis of C2 domain residues involved in membrane penetration demonstrate that membrane insertion by the C2 domain is required for membrane deformation, suggesting that C2 domain-induced membrane structural changes may be an important step in signaling pathways mediated by cPLA(2)α.
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame, South Bend, IN 46556, USA.
authors.forename: Katherine E
authors.initials: KE
authors.lastname: Ward
authors.forename: James P
authors.initials: JP
authors.lastname: Ropa
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Adu-Gyamfi
authors.forename: Robert V
authors.initials: RV
authors.lastname: Stahelin
mesh_terms: D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism*; D003001:Cloning, Molecular; D054513:Group IV Phospholipases A2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008563:Membrane Lipids / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D017434:Protein Structure, Tertiary
chemical_list: D008563:Membrane Lipids; D054513:Group IV Phospholipases A2; D002118:Calcium