SNARES in opposing bilayers interact in a circular array to form conducting pores
SNARES in opposing bilayers interact in a circular array to form conducting pores
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DOI：
10.1016/s0006-3495(02)75263-6
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发表时间：
2002-11-01
影响因子：
3.4
通讯作者：
Jena, BP
中科院分区：
文献类型：
--
作者：
Cho, SJ;Kelly, M;Jena, BP
神经末梢的融合过程是通过突触囊泡和突触前膜中的一组特殊蛋白质介导的。三种可溶性n -乙基马来酰亚胺敏感因子（NSF）附着蛋白受体（SNAREs）参与了膜融合。用原子力显微镜观察了这些与脂质双层相关的SNAREs的结构和排列。双层电生理装置允许测量膜电导和电容。在这里，我们证明了这些蛋白质形成融合孔的相互作用取决于t- snare和v-SNARE在对立双层中的存在。将纯化的重组v-SNARE添加到t-SNARE重构的脂质膜中，只增加了球形t-SNARE低聚物的大小，而不影响膜的电学性质。然而，当将t-SNARE囊泡添加到v-SNARE膜中时，可以观察到SNAREs以环形模式组装，并且电容逐步增加，电导增加。因此，t- snare和v-SNARE需要驻留在对立的双层中，以允许适当的t-/v-SNARE相互作用导致膜融合。
The process of fusion at the nerve terminal is mediated via a specialized set of proteins in the synaptic vesicles and the presynaptic membrane. Three soluble N-ethylmaleimide-sensitive factor (NSF)-attachment protein receptors (SNAREs) have been implicated in membrane fusion. The structure and arrangement of these SNAREs associated with lipid bilayers were examined using atomic force microscopy. A bilayer electrophysiological setup allowed for measurements of membrane conductance and capacitance. Here we demonstrate that the interaction of these proteins to form a fusion pore is dependent on the presence of t-SNAREs and v-SNARE in opposing bilayers. Addition of purified recombinant v-SNARE to a t-SNARE-reconstituted lipid membrane increased only the size of the globular t-SNARE oligomer without influencing the electrical properties of the membrane. However when t-SNARE vesicles were added to a v-SNARE membrane, SNAREs assembles in a ring pattern and a stepwise increase in capacitance, and increase in conductance were observed. Thus, t- and v-SNAREs are required to reside in opposing bilayers to allow appropriate t-/v-SNARE interactions leading to membrane fusion.