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

Abstract

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.

TRIM67 regulates exocytic mode and neuronal morphogenesis via SNAP47

“…In astrocytes, expression of a mutant lacking the majority of the transmembrane domain (VAMP2 1–96 ; Figure S3A ) in the presence of endogenous VAMP2 was shown by capacitance measurements to stabilize narrow fusion pores and block full fusion but not transient KNR ( Guček et al, 2016 ). Although lacking the majority of the transmembrane domain, VAMP2 1–96 continues to fractionate and associate with membranes ( Caccin et al, 2003 ; Wang et al, 2020 ). To validate classification and determine if fusion pore behavior distinguished modes, we expressed tagRFP-VAMP2 1–96 with full-length VAMP2-pHluorin, which is capable of and reports fusion.…”

Proteomics Study Reveals the Anti-Depressive Mechanisms and the Compatibility Advantage of Chaihu-Shugan-San in a Rat Model of Chronic Unpredictable Mild Stress

“…Vamp2, a core soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein residing on synaptic vesicles, forms helix bundles with SNAP25 and syntaxin-1 for the SNARE assembly ( Ramakrishnan et al, 2012 ; Wang et al, 2020 ). SNARE proteins were the main neurotransmitter release-associated proteins involving in the release procedure and synaptic vesicle fusion ( Kiessling et al, 2013 ; Stratton et al, 2016 ).…”

Cholesterol-enriched membrane micro-domain deficiency induces doxorubicin resistance via promoting autophagy in breast cancer

“… 71 , 77 , 78 , 79 With the exception of localization of SNAREs, the function of SNAREs is also known to associate with CEMMs, although how the functions of the SNAREs are regulated by CEMMs remains largely elusive and often controversial. 71 , 77 , 78 , 79 , 80 , 81 On one hand, the enriched SNAREs in CEMMs have been found to be required for efficient fusion events during endocytosis, indicating a positive role of CEMM in promoting SNAREs activity. 82 , 83 , 84 On the other hand, CEMM may suppress the autophagosome-lysosome fusion process mediated by SNAREs.…”