Clustering of Syntaxin-1A in Model Membranes Is Modulated by Phosphatidylinositol 4,5-Bisphosphate and Cholesterol

被引:90
作者
Murray, David H.
Tamm, Lukas K. [1 ]
机构
[1] Univ Virginia, Ctr Membrane Biol, Charlottesville, VA 22908 USA
关键词
GIANT UNILAMELLAR VESICLES; PLASMA-MEMBRANE; SNARE PROTEINS; FUSION; EXOCYTOSIS; SYNAPTOTAGMIN; DOMAINS; CORE; BISPHOSPHATE; HEMIFUSION;
D O I
10.1021/bi9003217
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Syntaxin-1A is part of the SNARE complex that forms in membrane fusion in neuronal exocytosis of synaptic vesicles. Together with SNAP-25 the single-span transmembrane protein syntaxin-1A forms the receptor complex on the plasma membrane of neuroendocrine cells. Previous studies have shown that syntaxin-1A occurs in Clusters that are different from lipid rafts in neuroendocrine plasma membranes. However, the interactions that promote these clusters have been largely unexplored. Here, we have reconstituted syntaxin-1A into lipid model membranes, and we show that syntaxin cluster formation depends on cholesterol in a lipid system that lacks sphingomyelin and therefore does not form liquid-ordered phases that are commonly believed to represent lipid rafts in cell membranes. Rather, the cholesterol-induced Clustering of syntaxin is found to be reversed by as little as 1-5 mol % of the regulatory lipid phosphatidylinositol 4,5-bisphosphate (PI-4,5-P-2), and PI-4,5-P-2 is shown to bind electrostatically to syntaxin, presumably mediated by the highly positively charged juxtamembrane domain of syntaxin. Possible implications of these results to the regulation of SNARE-mediated membrane fusion are discussed.
引用
收藏
页码:4617 / 4625
页数:9
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