title: Synaptotagmin perturbs the structure of phospholipid bilayers.
abstract: Synaptotagmin I (syt), an integral protein of the synaptic vesicle membrane, is believed to act as a Ca2+ sensor for neuronal exocytosis. Syt's cytoplasmic domain consists largely of two C2 domains, C2A and C2B. In response to Ca2+ binding, the C2 domains interact with membranes, becoming partially embedded in the lipid bilayer. We have imaged syt C2AB in association with lipid bilayers under fluid, using AFM. As expected, binding of C2AB to bilayers required both an anionic phospholipid [phosphatidylserine (PS)] and Ca2+. C2AB associated with bilayers in the form of aggregates of varying stoichiometries, and aggregate size increased with an increase in PS content. Repeated scanning of bilayers revealed that as C2AB dissociated it left behind residual indentations in the bilayer. The mean depth of these identations was 1.81 nm, indicating that they did not span the bilayer. Individual C2 domains (C2A and C2B) also formed aggregates and produced bilayer indentations. Binding of C2AB to bilayers and the formation of indentations were significantly compromised by mutations that interfere with binding of Ca2+ to syt or reduce the positive charge on the surface of C2B. We propose that bilayer perturbation by syt might be significant with respect to its ability to promote membrane fusion.
authors.affiliation: Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, United Kingdom.
authors.forename: Victor
authors.initials: V
authors.lastname: Shahin
authors.forename: Debajyoti
authors.initials: D
authors.lastname: Datta
authors.forename: Enfu
authors.initials: E
authors.lastname: Hui
authors.forename: Robert M
authors.initials: RM
authors.lastname: Henderson
authors.forename: Edwin R
authors.initials: ER
authors.lastname: Chapman
authors.forename: J Michael
authors.initials: JM
authors.lastname: Edwardson
mesh_terms: D018076:DNA, Complementary; D008051:Lipid Bilayers; D018625:Microscopy, Atomic Force; D015394:Molecular Structure; D009154:Mutation; D050863:Synaptotagmin I / Q000502:physiology*
chemical_list: D018076:DNA, Complementary; D008051:Lipid Bilayers; D050863:Synaptotagmin I