Tandem MS analysis of brain clathrin-coated vesicles reveals their critical involvement in synaptic vesicle recycling

• DOI: 10.1073/pnas.0308186101
• Publication Date: 2004-03-16T17:57:51Z
• Authors: Francois Blondeau; Brigitte Ritter; Patrick D. Allaire; Sylwia Wasiak; Martine Girard; Natasha K. Hussain; Annie Angers; Valerie Legendre-...; Line Roy; Daniel Boismenu; Robert E. Kearney; Alexander W. Bell; John J. M. Bergeron; Peter S. McPherson
• Keywords: Clathrin adaptor proteins; Organelle

Tandem MS has identified 209 proteins of clathrin-coated vesicles (CCVs) isolated from rat brain. An overwhelming abundance peptides were assigned to the clathrin coat with a 1:1 stoichiometry observed for heavy and light chains 2:1 chain adaptor protein heterotetramers. Thirty-two representing many known components synaptic (SVs) identified, supporting that main function brain CCVs is recapture SVs after exocytosis. A ratio vesicle- N -ethylmaleimide-sensitive factor attachment receptors target- receptors, similar previously detected on SVs, supports single-step model SV sorting during CCV-mediated recycling SVs. The uncovering eight undescribed proteins, four which have date been linked clathrin-mediated trafficking, further attests value current organelle-based proteomics strategy.