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Presynaptic calmodulin targets: Lessons from structural proteomics

Lipstein, N.; Göth, M.; Piotrowski, C.; Pagel, K.; Sinz, A.; Jahn, O.* – 2017

Calmodulin (CaM) is a highly conserved Ca2+-binding protein that is exceptionally abundant in the brain. In the presynaptic compartment of neurons, CaM transduces changes in Ca2+ concentration into the regulation of synaptic transmission dynamics. We review selected literature including published CaM interactor screens and outline established and candidate presynaptic CaM targets. We present a workflow of biochemical and structural proteomic methods that were used to identify and characterize the interactions between CaM and Munc13 proteins. Finally, we outline the potential of ion mobility-mass spectrometry (IM-MS) for conformational screening and of protein-protein cross-linking for the structural characterization of CaM complexes. Cross-linking/MS and native MS can be applied with considerable throughput to protein mixtures under near-physiological conditions, and thus effectively complement high-resolution structural biology techniques. Experimental distance constraints are applicable best when obtained by combining different cross-linking strategies, i.e. by using cross-linkers with different spacer length and reactivity, and by using the incorporation of unnatural photo-reactive amino acids. Insights from structural proteomics can be used to generate CaM-insensitive mutants of CaM-targets for functional studies in vitro or ideally in vivo.

Title
Presynaptic calmodulin targets: Lessons from structural proteomics
Author
Lipstein, N.; Göth, M.; Piotrowski, C.; Pagel, K.; Sinz, A.; Jahn, O.*
Date
2017
Identifier
10.1080/14789450.2017.1275966
Source(s)
Citation
Exp. Rev. Prot. 2017, 14, 223-242
Type
Text
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