Cited by 320Advanced Proteomics Techniques and ApplicationsNeuroscience and Neuropharmacology ResearchProtein Kinase Regulation and GTPase Signaling

Proteomic Analysis of in Vivo Phosphorylated Synaptic Proteins

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Collins, Mark O., Yu, Lu, Coba, Marcelo P., Husi, Holger, Campuzano, Iain, Blackstock, Walter P., Choudhary, Jyoti S., Grant, Seth G.N.·2005·Journal of Biological Chemistry

Abstract

In the nervous system, protein phosphorylation is an essential feature of synaptic function. Although protein phosphorylation is known to be important for many synaptic processes and in disease, little is known about global phosphorylation of synaptic proteins. Heterogeneity and low abundance make protein phosphorylation analysis difficult, particularly for mammalian tissue samples. Using a new approach, combining both protein and peptide immobilized metal affinity chromatography and mass spectrometry data acquisition strategies, we have produced the first large scale map of the mouse synapse phosphoproteome. We report over 650 phosphorylation events corresponding to 331 sites (289 have been unambiguously assigned), 92% of which are novel. These represent 79 proteins, half of which are novel phosphoproteins, and include several highly phosphorylated proteins such as MAP1B (33 sites) and Bassoon (30 sites). An additional 149 candidate phosphoproteins were identified by profiling the composition of the protein immobilized metal affinity chromatography enrichment. All major synaptic protein classes were observed, including components of important pre- and postsynaptic complexes as well as low abundance signaling proteins. Bioinformatic and in vitro phosphorylation assays of peptide arrays suggest that a small number of kinases phosphorylate many proteins and that each substrate is phosphorylated by many kinases. These data substantially increase existing knowledge of synapse pro…

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Crossref · 10.1074/jbc.m411220200

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  • Although it is well established that CaMKII phosphorylates numerous synaptic proteins, an essential role for CaMKII phosphorylation of many of these targets largely remains elusive.

    “It should be pointed out that, although it is well established that CaMKII phosphorylates numerous synaptic proteins (66, 434–437), as reviewed here, despite intensive research an essential role for CaMKII phosphorylation of many of these targets largely remains elusive.”

    From Synaptic memory and CaMKII (2023) · cites this paper as [436]

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Authors
Collins, M. O., Yu, L., Coba, M. P., Husi, H., Campuzano, I., Blackstock, W. P., Choudhary, J. S., & Grant, S. G. N.
Affiliations
  • Wellcome Sanger Institute (GB)
  • University of Edinburgh (GB)
  • Waters (United Kingdom) (GB)
  • University of Sheffield (GB)
Corresponding author
Grant, Seth G.N.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2005-02
Volume
280
Issue
7
Pages
5972-5982
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2167791125
Cited by
320
References
43
Citations by year
2026: 2 · 2025: 2 · 2024: 2 · 2023: 7 · 2022: 4 · 2021: 11 · 2020: 6 · 2019: 11 · 2018: 9 · 2017: 8 · 2016: 12 · 2015: 8 · 2014: 11 · 2013: 9 · 2012: 17
FWCI
14.96
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords
synaptic proteins, protein phosphorylation, immobilized metal affinity chromatography, MAP1B, bassoon, kinase substrate specificity
MeSH terms
Animals, Cell Membrane, Phosphoproteins, Phosphorylation, Protein Binding, Protein Kinases, Solubility, Synapses, Synaptosomes, Urea, Computational Biology, Protein Array Analysis, Proteomics, Mice

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