title: Characterization of the Munc13-calmodulin interaction by photoaffinity labeling.
abstract: Sensing of and response to transient increases in the residual presynaptic Ca2+ levels are important adaptive mechanisms that define the short-term plasticity characteristics of neurons. Due to their essential function in synaptic vesicle priming and in the modulation of synaptic strength, Munc13 proteins have emerged as key regulators of these adaptive mechanisms. Indeed, Munc13-1 and ubMunc13-2 contain a conserved calmodulin (CaM) binding site and the Ca2+ -dependent interaction of these Munc13 isoforms with CaM constitutes a molecular mechanism that transduces residual Ca2+ signaling to the synaptic exocytotic machinery. Here, we used Munc13-derived model peptides in photoaffinity labeling (PAL) experiments to demonstrate the stoichiometric and Ca2+ -dependent CaM binding of the other members of the Munc13 family, bMunc13-2 and Munc13-3, via structurally distinct non-conserved binding sites. A PAL-based Ca2+ titration assay revealed that all Munc13 isoforms can form a complex with CaM already at low Ca2+ concentrations just above resting levels, underscoring the Ca2+ sensor/effector function of this interaction in short-term synaptic plasticity phenomena.
authors.affiliation: Proteomics Group, Max-Planck-Institute of Experimental Medicine, Hermann-Rein-Strasse 3, 37075 Göttingen, Germany.
authors.forename: Kalina
authors.initials: K
authors.lastname: Dimova
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kawabe
authors.forename: Andrea
authors.initials: A
authors.lastname: Betz
authors.forename: Nils
authors.initials: N
authors.lastname: Brose
authors.forename: Olaf
authors.initials: O
authors.lastname: Jahn
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000737:chemistry; D002147:Calmodulin / Q000737:chemistry*; D002417:Cattle; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D019920:Photoaffinity Labels / Q000737:chemistry*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary
chemical_list: D002147:Calmodulin; D009419:Nerve Tissue Proteins; D010455:Peptides; D019920:Photoaffinity Labels; D002118:Calcium