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http://dx.doi.org/10.5012/bkcs.2007.28.9.1499

A Comprehensive Identification of Synaptic Vesicle Proteins in Rat Brains by cRPLC/MS-MS and 2DE/MALDI-TOF-MS  

Lee, Won-Kyu (Division of Life Sciences, Korea Institute of Science and Technology)
Kim, Hye-Jung (Division of Life Sciences, Korea Institute of Science and Technology)
Min, Hye-Ki (Department of Chemistry, Korea University)
Kang, Un-Beom (Division of Life Sciences, Korea Institute of Science and Technology)
Lee, Cheol-Ju (Division of Life Sciences, Korea Institute of Science and Technology)
Lee, Sang-Won (Department of Chemistry, Korea University)
Kim, Ick-Young (Laboratory of Cellular and Molecular Biochemistry, School of Life Sciences and Biotechnology, Korea University)
Lee, Seung-Taek (Department of Biochemistry, College of Science, Yonsei University)
Kwon, Oh-Seung (Division of Life Sciences, Korea Institute of Science and Technology)
Yu, Yeon-Gyu (Department of Chemistry, Kookmin University)
Publication Information
Bulletin of the Korean Chemical Society / v.28, no.9, 2007 , pp. 1499-1509 More about this Journal
Abstract
Proteomic analyses of synaptic vesicle fraction from rat brain have been performed for the better understanding of vesicle regulation and signal transmission. Two different approaches were applied to identify proteins in synaptic vesicle fraction. First, the isolated synaptic vesicle proteins were treated with trypsin, and the resulting peptides were analyzed using a high-pressure capillary reversed phase liquid chromatography/tandem mass spectrometry (cRPLC/MS/MS). Alternatively, proteins were separated by two-dimensional gel electrophoresis (2DE) and identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF/MS). Total 18 and 52 proteins were identified from cRPLC/MS-MS and 2DE-MALDI-TOF-MS analysis, respectively. Among them only 2 proteins were identified by both methods. Of the proteins identified, 70% were soluble proteins and 30% were membrane proteins. They were categorized by their functions in vesicle trafficking and biogenesis, energy metabolism, signal transduction, transport and unknown functions. Among them, 27 proteins were not previously reported as synaptic proteins. The cellular functions of unknown proteins were estimated from the analysis of domain structure, expression profile and predicted interaction partners.
Keywords
Synaptic membrane proteome; Micro liquid chromatography electrospray ionization tandem mass spectrometry; Two-dimensional gel electrophoresis; Matrix-assisted laser desorption mass spectrometry; Rat brains;
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