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PMID: 19251653 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

A scissors mechanism for stimulation of SNARE-mediated lipid mixing by cholesterol.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 106 ·No. 13 ·2009-03-31 ·Pages 5141-6

Tong J, Borbat PP, Freed JH, Shin YK

Abstract

Neurotransmitter release at the synapse requires membrane fusion. The SNARE complex, composed of the plasma membrane t-SNAREs syntaxin 1A and SNAP-25 and the vesicle v-SNARE synaptobrevin, mediates the fusion of 2 membranes. Synaptic vesicles contain unusually high cholesterol, but the exact role of cholesterol in fusion is not known. In this study, cholesterol was found to stimulate SNARE-mediated lipid mixing of proteoliposomes by a factor of 5 at a physiological concentration. Surprisingly, however, the stimulatory effect was more pronounced when cholesterol was on the v-SNARE side than when it was on the t-SNARE side. Site-directed spin labeling and both continuous wave (CW) and pulsed EPR revealed that cholesterol induces a conformational change of the v-SNARE transmembrane domain (TMD) from an open scissors-like dimer to a parallel dimer. When the TMD was forced to form a parallel dimer by the disulfide bond, the rate was stimulated 2.3-fold even without cholesterol, supporting the relevance of the open-to-closed conformational change to the fusion activity. The open scissors-like conformation may be unfavorable for fusion and cholesterol may relieve this inhibitory factor.

MeSH Terms
Cholesterol/metabolism Electron Spin Resonance Spectroscopy Humans Membrane Fusion Membrane Lipids/metabolism Protein Conformation SNARE Proteins/metabolism,physiology
Chemicals
Membrane Lipids SNARE Proteins Cholesterol
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Tong Jiansong
Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, IA 50011, USA.
Borbat Peter P
Freed Jack H
Shin Yeon-Kyun
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
DOI
10.1073/pnas.0813138106
Published
2009-03-31
Epub
2009-00-27
Pages
5141-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2663986
Subset
IM
Grants
PHS HHS · R01-ER03150 · United States
NIBIB NIH HHS · R01 EB003150 · United States
PHS HHS · R01-51290 · United States
NCRR NIH HHS · P41 RR016292 · United States
NIBIB NIH HHS · R01 EB003150-26 · United States
NCRR NIH HHS · P41-RR016292 · United States
NIBIB NIH HHS · R01 EB003150-27 · United States
NCRR NIH HHS · P41 RR016292-08 · United States
External Links
PubMed source DOI: 10.1073/pnas.0813138106 PMC Full Text View journal details
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