Protein Phosphorylation in Depolarized Synaptosomes: Dissecting Primary Effects of Calcium from Synaptic Vesicle Cycling

2021 | journal article; research paper. A publication with affiliation to the University of Göttingen.

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​Protein Phosphorylation in Depolarized Synaptosomes: Dissecting Primary Effects of Calcium from Synaptic Vesicle Cycling​
Silbern, I.; Pan, K.-T.; Fiosins, M.; Bonn, S. ; Rizzoli, S. O. ; Fornasiero, E. F.   & Urlaub, H.  et al.​ (2021) 
Molecular & Cellular Proteomics20 pp. 100061​.​ DOI: https://doi.org/10.1016/j.mcpro.2021.100061 

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Authors
Silbern, Ivan; Pan, Kuan-Ting; Fiosins, Maksims; Bonn, Stefan ; Rizzoli, Silvio O. ; Fornasiero, Eugenio Francesco ; Urlaub, Henning ; Jahn, Reinhard 
Abstract
Synaptic transmission is mediated by the regulated exocytosis of synaptic vesicles. When the presynaptic membrane is depolarized by an incoming action potential, voltage-gated calcium channels open, resulting in the influx of calcium ions that triggers the fusion of synaptic vesicles (SVs) with the plasma membrane. SVs are recycled by endocytosis. Phosphorylation of synaptic proteins plays a major role in these processes, and several studies have shown that the synaptic phosphoproteome changes rapidly in response to depolarization. However, it is unclear which of these changes are directly linked to SV cycling and which might regulate other presynaptic functions that are also controlled by calcium-dependent kinases and phosphatases. To address this question, we analyzed changes in the phosphoproteome using rat synaptosomes in which exocytosis was blocked with botulinum neurotoxins (BoNTs) while depolarization-induced calcium influx remained unchanged. BoNT-treatment significantly alters the response of the synaptic phoshoproteome to depolarization and results in reduced phosphorylation levels when compared with stimulation of synaptosomes by depolarization with KCl alone. We dissect the primary Ca2+-dependent phosphorylation from SV-cycling-dependent phosphorylation and confirm an effect of such SV-cycling-dependent phosphorylation events on syntaxin-1a-T21/T23, synaptobrevin-S75, and cannabinoid receptor-1-S314/T322 on exo- and endocytosis in cultured hippocampal neurons.
Issue Date
2021
Journal
Molecular & Cellular Proteomics 
Project
EXC 2067: Multiscale Bioimaging 
SFB 1286: Quantitative Synaptologie 
SFB 1286 | A03: Dynamische Analyse der Remodellierung der extrazellulären Matrix (ECM) als Mechanismus der Synapsenorganisation und Plastizität 
SFB 1286 | A08: Die Rolle post-translational modifizierter Proteine in der synaptischen Übertragung 
SFB 1286 | Z02: Integrative Datenanalyse und -interpretation. Generierung einer synaptisch-integrativen Datenstrategie (SynIDs) 
SFB 1286 | Z03: Unkomplizierte multispektrale, superauflösende Bildgebung durch zehnfache Expansionsmikroskopie 
Working Group
RG Rizzoli (Quantitative Synaptology in Space and Time) 
RG Bonn 
RG R. Jahn (Laboratory of Neurobiology) 
RG Urlaub (Bioanalytische Massenspektrometrie) 
ISSN
1535-9476
Language
English

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