title: Membrane Docking of the Synaptotagmin 7 C2A Domain: Electron Paramagnetic Resonance Measurements Show Contributions from Two Membrane Binding Loops.
abstract: The synaptotagmin (Syt) family of proteins plays an important role in vesicle docking and fusion during Ca(2+)-induced exocytosis in a wide variety of cell types. Its role as a Ca(2+) sensor derives primarily from its two C2 domains, C2A and C2B, which insert into anionic lipid membranes upon binding Ca(2+). Syt isoforms 1 and 7 differ significantly in their Ca(2+) sensitivity; the C2A domain from Syt7 binds Ca(2+) and membranes much more tightly than the C2A domain from Syt1, at least in part because of greater contributions from the hydrophobic effect. While the structure and membrane activity of Syt1 have been extensively studied, the structural origins of differences between Syt1 and Syt7 are unknown. This study used site-directed spin labeling and electron paramagnetic resonance spectroscopy to determine depth parameters for the Syt7 C2A domain, for comparison to analogous previous measurements with the Syt1 C2A domain. In a novel approach, the membrane docking geometry of both Syt1 and Syt7 C2A was modeled by mapping depth parameters onto multiple molecular dynamics-simulated structures of the Ca(2+)-bound protein. The models reveal membrane penetration of Ca(2+) binding loops 1 (CBL1) and 3 (CBL3), and membrane binding is more sensitive to mutations in CBL3. On average, Syt7 C2A inserts more deeply into the membrane than Syt1 C2A, although depths vary among the different structural models. This observation provides a partial structural explanation for the hydrophobically driven membrane docking of Syt7 C2A.
authors: Department of Chemistry, University of Colorado Denver , Denver, Colorado 80217, United States.
authors: J Ryan
authors: JR
authors: Osterberg
authors: Department of Chemistry, University of Colorado Denver , Denver, Colorado 80217, United States.
authors: Nara Lee
authors: NL
authors: Chon
authors: Department of Chemistry, University of Colorado Denver , Denver, Colorado 80217, United States.
authors: Arthur
authors: A
authors: Boo
authors: Department of Chemistry, University of Colorado Denver , Denver, Colorado 80217, United States.
authors: Favinn A
authors: FA
authors: Maynard
authors: Department of Chemistry, University of Colorado Denver , Denver, Colorado 80217, United States.
authors: Hai
authors: H
authors: Lin
authors: Department of Chemistry, University of Colorado Denver , Denver, Colorado 80217, United States.
authors: Jefferson D
authors: JD
authors: Knight
mesh_terms: D001665:Binding Sites; D004578:Electron Spin Resonance Spectroscopy; D007700:Kinetics; D062105:Molecular Docking Simulation; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D050857:Synaptotagmins / Q000737:chemistry* / Q000235:genetics; D053835:Unilamellar Liposomes / Q000737:chemistry*
chemical_list: D053835:Unilamellar Liposomes; D050857:Synaptotagmins