Open AccessTandem MS analysis of brain clathrin-coated vesicles reveals their critical involvement in synaptic vesicle recycling
Author(s) -
F Blondeau,
Brigitte Ritter,
Patrick D. Allaire,
Sylwia Wasiak,
Martine Girard,
Natasha K. Hussain,
Annie Angers,
Valérie Legendre-Guillemin,
Line Roy,
Daniel Boismenu,
Robert E. Kearney,
Alexander W. Bell,
John J.M. Bergeron,
Peter S. McPherson
Publication year - 2004
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.0308186101
Subject(s) - clathrin , vesicle , clathrin adaptor proteins , synaptic vesicle , microbiology and biotechnology , exocytosis , signal transducing adaptor protein , biology , organelle , proteomics , chemistry , biochemistry , membrane , signal transduction , gene
Tandem MS has identified 209 proteins of clathrin-coated vesicles (CCVs) isolated from rat brain. An overwhelming abundance of peptides were assigned to the clathrin coat with a 1:1 stoichiometry observed for clathrin heavy and light chains and a 2:1 stoichiometry of clathrin heavy chain with clathrin adaptor protein heterotetramers. Thirty-two proteins representing many of the known components of synaptic vesicles (SVs) were identified, supporting that a main function for brain CCVs is to recapture SVs after exocytosis. A ratio of vesicle-N-ethylmaleimide-sensitive factor attachment protein receptors to target-N-ethylmaleimide-sensitive factor attachment protein receptors, similar to that previously detected on SVs, supports a single-step model for SV sorting during CCV-mediated recycling of SVs. The uncovering of eight previously undescribed proteins, four of which have to date been linked to clathrin-mediated trafficking, further attests to the value of the current organelle-based proteomics strategy.