Analysis of the synaptic vesicle proteome using three gel-based protein separation techniques

Jacqueline Burré, Tobias Beckhaus, Hermann Schägger, Carsten Corvey, Sandra Hofmann, Michael Karas, Herbert Zimmermann, Walter Volknandt

Research output: Contribution to journalArticle

79 Scopus citations

Abstract

Synaptic vesicles are key organelles in neurotransmission. Their functions are governed by a unique set of integral and peripherally associated proteins. To obtain a complete protein inventory, we immunoisolated synaptic vesicles from rat brain to high purity and performed a gel-based analysis of the synaptic vesicle proteome. Since the high hydrophobicity of integral membrane proteins hampers their resolution by gel electrophoretic techniques, we applied in parallel three different gel electrophoretic methods for protein separation prior to MS. Synaptic vesicle proteins were subjected to either 1-D SDS-PAGE along with nano-LC ESI-MS/MS or to the 2-D gel electrophoretic techniques benzyldimethyl-n-hexadecylammonium chloride (BAC)/SDS-PAGE, and double SDS (dSDS)-PAGE in combination with MALDI-TOF-MS. We demonstrate that the combination of all three methods provides a comprehensive survey of the proteinaceous inventory of the synaptic vesicle membrane compartment. The identified synaptic vesicle proteins include transporters, soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), synapsins, rab and rab-interacting proteins, additional guanine nucleotide triphosphate (GTP) binding proteins, cytoskeletal proteins, and proteins modulating synaptic vesicle exo- and endocytosis. In addition, we identified novel proteins of unknown function. Our results demonstrate that the parallel application of three different gel-based approaches in combination with mass spectrometry permits a comprehensive analysis of the synaptic vesicle proteome that is considerably more complex than previously anticipated.

Original languageEnglish (US)
Pages (from-to)6250-6262
Number of pages13
JournalProteomics
Volume6
Issue number23
DOIs
StatePublished - Dec 1 2006

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Keywords

  • Mass spectrometry
  • Membrane protein
  • Proteome
  • Synaptic vesicle protein
  • Two-dimensional electrophoresis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Burré, J., Beckhaus, T., Schägger, H., Corvey, C., Hofmann, S., Karas, M., Zimmermann, H., & Volknandt, W. (2006). Analysis of the synaptic vesicle proteome using three gel-based protein separation techniques. Proteomics, 6(23), 6250-6262. https://doi.org/10.1002/pmic.200600357