title: Protein quantification at the single vesicle level reveals that a subset of synaptic vesicle proteins are trafficked with high precision.
abstract: Protein sorting represents a potential point of regulation in neurotransmission because it dictates the protein composition of synaptic vesicles, the organelle that mediates transmitter release. Although the average number of most vesicle proteins has been estimated using bulk biochemical approaches (Takamori et al., 2006), no information exists on the intervesicle variability of protein number, and thus on the precision with which proteins are sorted to vesicles. To address this, we adapted a single molecule quantification approach (Mutch et al., 2007) and used it to quantify both the average number and variance of seven integral membrane proteins in brain synaptic vesicles. We report that four vesicle proteins, SV2, the proton ATPase, Vglut1, and synaptotagmin 1, showed little intervesicle variation in number, indicating they are sorted to vesicles with high precision. In contrast, the apparent number of VAMP2/synaptobrevin 2, synaptophysin, and synaptogyrin demonstrated significant intervesicle variability. These findings place constraints on models of protein function at the synapse and raise the possibility that changes in vesicle protein expression affect vesicle composition and functioning.
authors.affiliation: Department of Chemistry and Pharmacology, University of Washington, Seattle, WA 98195, USA.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Mutch
authors.forename: Patricia
authors.initials: P
authors.lastname: Kensel-Hammes
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: Gadd
authors.forename: Bryant S
authors.initials: BS
authors.lastname: Fujimoto
authors.forename: Richard W
authors.initials: RW
authors.lastname: Allen
authors.forename: Perry G
authors.initials: PG
authors.lastname: Schiro
authors.forename: Robert M
authors.initials: RM
authors.lastname: Lorenz
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Kuyper
authors.forename: Jason S
authors.initials: JS
authors.lastname: Kuo
authors.forename: Sandra M
authors.initials: SM
authors.lastname: Bajjalieh
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Chiu
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000378:metabolism*; D021381:Protein Transport; D051381:Rats; D017207:Rats, Sprague-Dawley; D013572:Synaptic Vesicles / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins