Imaging the assembly and disassembly kinetics of cis-SNARE complexes on native plasma membranes
Imaging the assembly and disassembly kinetics of cis-SNARE complexes on native plasma membranes
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DOI：
10.1016/j.febslet.2008.08.040
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发表时间：
2008-10-15
期刊：
影响因子：
3.5
通讯作者：
Ashery, Uri
中科院分区：
文献类型：
--
作者：
Bar-On, Dana;Winter, Ulrike;Ashery, Uri
真核细胞的温和超声处理产生保留其对接的细胞器、细胞骨架结构和细胞质复合物的天然质膜片。虽然膜蛋白复合物的精细组织保持不受干扰，但它们的内质膜小叶可以迅速暴露于沐浴溶液，从而实现特定的生化操作。在这里，我们应用这个系统来跟踪膜生化动力学。我们监测可溶性NSF附着蛋白受体（SNARE）复杂的组装和拆卸质膜上的高时间分辨率。结果表明，两个阶段的组装过程和拆卸动力学的N-乙基马来酰亚胺敏感因子（NSF）和可溶性NSF附着蛋白（α-SNAP）浓度的依赖动力学动力学。(C)2008年欧洲生物化学学会联合会。由Elsevier B出版。V.保留所有权利。
Mild sonication of eukaryotic cells produces native plasma membrane sheets that retain their docked organelles, cytoskeleton structures and cytoplasmic complexes. While the delicate organization of membranous protein complexes remains undisturbed, their inner plasmalemmel leaflet can be rapidly exposed to bathing solutions, enabling specific biochemical manipulations. Here, we apply this system to track membrane-biochemistry kinetics. We monitor soluble NSF-attachment protein receptor (SNARE) complex assembly and disassembly on the plasma membrane at high time resolution. The results suggest two-phase kinetics for the assembly process and dependence of the disassembly kinetics on both N-ethyl maleimide-sensitive factor (NSF) and soluble NSF-attachment protein (alpha-SNAP) concentrations. (C) 2008 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.