Single-molecule FRET-derived model of the synaptotagmin 1-SNARE fusion complex.

Single-molecule FRET-derived model of the synaptotagmin 1-SNARE fusion complex.
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DOI:
10.1038/nsmb.1763
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发表时间:
2010-03
影响因子:
16.8
通讯作者:
--
中科院分区:
生物学1区
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当钙离子与突触蛋白1结合时,同步神经传递被触发。突触蛋白1是一种与SNARS和膜相互作用的突触囊泡蛋白。我们使用synaptopagmin的两个C2结构域之间的单分子FRET来确定它们的构象由多个偶尔发生转变的状态组成,与结构域的随机相对运动一致。SNARE结合导致突触内FRET分布更窄,状态之间的转换频率更低。我们使用多体对接的方法,以34个FRET衍生的距离为约束,获得了一个实验确定的难以捉摸的突触素1-SNARE复合体的模型。钙结合环指向远离SNARE复合体,因此它们可以与同一膜相互作用。这种环的排列类似于SNARE诱导的钙结合突触素3的晶体结构,提示了突触与SNARS之间的相互作用在钙触发的融合中发挥作用的共同机制。
Synchronous neurotransmission is triggered when Ca2+ binds to synaptotagmin 1, a synaptic vesicle protein that interacts with SNAREs and membranes. We used single-molecule FRET between synaptotagmin’s two C2 domains to determine that their conformation consists of multiple states with occasional transitions, consistent with domains in random relative motion. SNARE binding results in narrower intra-synaptotagmin FRET distributions and less frequent transitions between states. We obtained an experimentally determined model of the elusive synaptotagmin 1–SNARE complex by using a multi-body docking approach with 34 FRET-derived distances as restraints. The Ca2+-binding loops point away from the SNARE complex, so they could interact with the same membrane. The loop arrangement is similar to that of the crystal structure of SNARE-induced Ca2+ bound synaptotagmin 3, suggesting a common mechanism by which the interaction between synaptotagmins and SNAREs plays a role in Ca2+-triggered fusion.
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