Mechanistic insights into the SNARE complex disassembly
Mechanistic insights into the SNARE complex disassembly
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SNARE 复杂拆卸的机制见解
DOI：
10.1126/sciadv.aau8164
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发表时间：
2019-04-01
期刊：
影响因子：
13.6
通讯作者：
Sui, Sen-Fang
中科院分区：
文献类型：
--
作者：
Huang, Xuan;Sun, Shan;Sui, Sen-Fang
NSF（N-乙基马来酰亚胺敏感因子）和α-SNAP（α-可溶性NSF附着蛋白）与SNARE（可溶性NSF附着蛋白受体）复合物（介导膜融合的最小机制）结合，形成20 S复合物，该复合物分解SNARE复合物以重新使用。我们报告的cryo-EM结构的α-SNAP-SNARE子复合物和NSF-D1 D2域的20 S复合物在3.9和3.7埃的分辨率，分别。结合生物化学和电生理学分析，我们发现α-SNAP主要通过静电相互作用的R116和疏水相互作用的L197对VAMP蛋白的两个位置施加作用力，从而完成解装配过程。此外，我们定义了SNARE螺旋束的氨基末端和NSF-D1结构域的孔环之间的相互作用，并证明了其作为SNARE复合物解体的潜在锚的重要作用。我们的研究提供了α-SNAP介导的SNARE复合物分解的旋转模型。
NSF (N-ethylmaleimide-sensitive factor) and alpha-SNAP (alpha-soluble NSF attachment protein) bind to the SNARE (soluble NSF attachment protein receptor) complex, the minimum machinery to mediate membrane fusion, to form a 20S complex, which disassembles the SNARE complex for reuse. We report the cryo-EM structures of the alpha-SNAP-SNARE subcomplex and the NSF-D1D2 domain in the 20S complex at 3.9- and 3.7-angstrom resolutions, respectively. Combined with the biochemical and electrophysiological analyses, we find that alpha-SNAPs use R116 through electrostatic interactions and L197 through hydrophobic interactions to apply force mainly on two positions of the VAMP protein to execute disassembly process. Furthermore, we define the interaction between the amino terminus of the SNARE helical bundle and the pore loop of the NSF-D1 domain and demonstrate its essential role as a potential anchor for SNARE complex disassembly. Our studies provide a rotation model of alpha-SNAP-mediated disassembly of the SNARE complex.