Tandem MS analysis of brain clathrin-coated vesicles reveals their critical involvement in synaptic vesicle recycling.
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.
Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, 3801 University Street, Montreal, QC, Canada H3A 2B4.
Francois
F
Blondeau
Brigitte
B
Ritter
Patrick D
PD
Allaire
Sylwia
S
Wasiak
Martine
M
Girard
Natasha K
NK
Hussain
Annie
A
Angers
Valerie
V
Legendre-Guillemin
Line
L
Roy
Daniel
D
Boismenu
Robert E
RE
Kearney
Alexander W
AW
Bell
John J M
JJ
Bergeron
Peter S
PS
McPherson
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002853:Chromatography, Liquid; D022163:Clathrin-Coated Vesicles / Q000737:chemistry* / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D013058:Mass Spectrometry; D051381:Rats; D013572:Synaptic Vesicles / Q000378:metabolism*