Delineating the Molecular Basis of the Calmodulin–bMunc13-2 Interaction by Cross-Linking/Mass Spectrometry—Evidence for a Novel CaM Binding Motif in bMunc13-2
Piotrowski, Christine
Moretti, Rocco
Ihling, Christian H.
Haedicke, Andre
Liepold, Thomas
Lipstein, Noa
Meiler, Jens
Jahn, Olaf
Sinz, Andrea
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2020-01
Abstract
Exploring the interactions between the Ca2+ binding protein calmodulin (CaM) and its target proteins remains a challenging task. Members of the Munc13 protein family play an essential role in short-term synaptic plasticity, modulated via the interaction with CaM at the presynaptic compartment. In this study, we focus on the bMunc13-2 isoform expressed in the brain, as strong changes in synaptic transmission were observed upon its mutagenesis or deletion. The CaM-bMunc13-2 interaction was previously characterized at the molecular level using short bMunc13-2-derived peptides only, revealing a classical 1-5-10 CaM binding motif. Using larger protein constructs, we have now identified for the first time a novel and unique CaM binding site in bMunc13-2 that contains an N-terminal extension of a classical 1-5-10 CaM binding motif. We characterize this motif using a range of biochemical and biophysical methods and highlight its importance for the CaM-bMunc13-2 interaction.