fusogenic pairings of vesicle-associated membrane proteins (vamps) and plasma membrane t-snares – vamp5 as the exceptionfusogenic配对的vesicle-associated膜蛋白(更新)和质膜t-snaresu2014u2014vamp5例外.pdfVIP

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fusogenic pairings of vesicle-associated membrane proteins (vamps) and plasma membrane t-snares – vamp5 as the exceptionfusogenic配对的vesicle-associated膜蛋白(更新)和质膜t-snaresu2014u2014vamp5例外.pdf

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fusogenic pairings of vesicle-associated membrane proteins (vamps) and plasma membrane t-snares – vamp5 as the exceptionfusogenic配对的vesicle-associated膜蛋白(更新)和质膜t-snaresu2014u2014vamp5例外

Fusogenic Pairings of Vesicle-Associated Membrane Proteins (VAMPs) and Plasma Membrane t-SNAREs – VAMP5 as the Exception Nazarul Hasan, Deborah Corbin, Chuan Hu* Department of Biochemistry and Molecular Biology, University of Louisville School of Medicine, Louisville, Kentucky, United States of America Abstract Background: Intracellular vesicle fusion is mediated by the interactions of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins on vesicles (v-SNAREs) and on target membranes (t-SNAREs). The vesicle-associated membrane proteins (VAMPs) are v-SNAREs that reside in various post-Golgi vesicular compartments. To fully understand the specific role of each VAMP in vesicle trafficking, it is important to determine if VAMPs have differential membrane fusion activities. Methodology/Principal Findings: In this study, we developed a cell fusion assay that quantifies SNARE-mediated membrane fusion events by activated expression of b-galactosidase, and examined fusogenic pairings between the seven VAMPs, i.e., VAMPs 1, 2, 3, 4, 5, 7 and 8, and two plasma membrane t-SNARE complexes, syntaxin1/SNAP-25 and syntaxin4/ SNAP-25. VAMPs 1, 2, 3, 4, 7 and 8 drove fusion efficiently, whereas VAMP5 was unable to mediate fusion with the t-SNAREs. By expressing VAMPs 1, 3, 4, 7 and 8 at the same level, we further compared their membrane fusion activities. VAMPs 1 and 3 had comparable and the highest fusion activities, whereas VAMPs 4, 7 and 8 exhibited 30–50% lower fusion activities. Moreover, we determined the dependence of cell fusion activity on VAMP1 expression level. Analysis of the dependence data suggested that there was no cooperativity of VAMP proteins in the cell fusion reaction. Conclusions/Significance: These data indicate that VAMPs have differential membrane fusion capacities, and imply that with