Different regions of synaptic vesicle membrane regulate VAMP2 conformation for the SNARE assembly

• Published in: Nature communications Vol. 11; no. 1; p. 1531
• Main Authors: Wang, Chuchu, Tu, Jia, Zhang, Shengnan, Cai, Bin, Liu, Zhenying, Hou, Shouqiao, Zhong, Qinglu, Hu, Xiao, Liu, Wenbin, Li, Guohui, Liu, Zhijun, He, Lin, Diao, Jiajie, Zhu, Zheng-Jiang, Li, Dan, Liu, Cong
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 24.03.2020; Nature Publishing Group UK; Nature Portfolio
• Subjects: Assembly; Cell Line, Tumor; Cholesterol; Cholesterol - metabolism; HEK293 Cells; Humans; Intravital Microscopy; Lipid Metabolism; Lipidomics; Lipids; Magnetic Resonance Spectroscopy; Mammalian cells; Mammals; Membrane Fusion; Membrane Lipids - metabolism; Membrane Microdomains - metabolism; Membrane proteins; Membranes; NMR; NMR spectroscopy; Nuclear magnetic resonance; Protein Domains - genetics; Protein Multimerization; Proteins; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; SNAP receptors; SNAP-25 protein; SNARE Proteins - metabolism; Spatial Analysis; Spectrometry; Spectroscopy; Synaptic vesicles; Synaptic Vesicles - metabolism; Syntaxin; Syntaxin 1; Vesicle-Associated Membrane Protein 2 - genetics; Vesicle-Associated Membrane Protein 2 - metabolism
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-15270-4

Vesicle associated membrane protein 2 (VAMP2/synaptobrevin2), a core SNARE protein residing on synaptic vesicles (SVs), forms helix bundles with syntaxin-1 and SNAP25 for the SNARE assembly. Prior to the SNARE assembly, the structure of VAMP2 is unclear. Here, by using in-cell NMR spectroscopy, we describe the dynamic membrane association of VAMP2 SNARE motif in mammalian cells, and the structural change of VAMP2 upon the change of intracellular lipid environment. We analyze the lipid compositions of the SV membrane by mass-spectrometry-based lipidomic profiling, and further reveal that VAMP2 forms distinctive conformations in different membrane regions. In contrast to the non-raft region, the membrane region of cholesterol-rich lipid raft markedly weakens the membrane association of VAMP2 SNARE motif, which releases the SNARE motif and facilitates the SNARE assembly. Our work reveals the regulation of different membrane regions on VAMP2 structure and sheds light on the spatial regulation of SNARE assembly.