The domain of brain beta-spectrin responsible for synaptic vesicle association is essential for synaptic transmission.
The domain of brain beta-spectrin responsible for synaptic vesicle association is essential for synaptic transmission.
复制标题
负责突触小泡关联的大脑β-血影蛋白域对于突触传递至关重要。
DOI：
10.1016/s0006-8993(00)02796-7
复制
发表时间：
2000
期刊：
影响因子：
2.9
通讯作者：
Goodman,SR
中科院分区：
文献类型：
--
作者：
Zimmer,WE;Zhao,Y;Sikorski,AF;Critz,SD;Sangerman,J;Elferink,LA;Xu,XS;Goodman,SR
我们研究了突触蛋白 I（小突触囊泡膜上的主要磷蛋白）与脑血影蛋白之间的相互作用。使用重组肽，我们将 β-血影蛋白亚型 βSpIIΣI 上的突触蛋白 I 附着位点定位到 25 个氨基酸（残基 211 至 235）的区域。该片段与肌动蛋白结合结构域相邻，并且位于 βSpIIΣI 的区域内，我们之前根据序列同源性预测该区域为候选突触蛋白 I 结合结构域。我们使用差速离心技术来定量评估血影蛋白与突触小泡的相互作用。使用该测定，观察到重组 βSpIIΣI 蛋白与突触小泡的高亲和力饱和结合。仅当重组肽上包含 25 个氨基酸突触蛋白 I 结合位点时才能观察到结合。此外，我们证明针对突触蛋白 I 结合域的 15 个氨基酸的抗体特异性阻断培养的海马神经元中的突触传递。因此，βSpIIΣI 血影蛋白上的突触蛋白 I 附着位点包含该分子的约 25 个氨基酸片段，这两种蛋白质的相互作用是神经传递过程的重要步骤。
We have examined the interaction between synapsin I, the major phosphoprotein on the membrane of small synaptic vesicles, and brain spectrin. Using recombinant peptides we have localized the synapsin I attachment site upon the β-spectrin isoform βSpIIΣI to a region of 25 amino acids, residues 211 through 235. This segment is adjacent to the actin binding domain and is within the region of the βSpIIΣI that we previously predicted as a candidate synapsin I binding domain based upon sequence homology. We used differential centrifugation techniques to quantitatively assess the interaction of spectrin with synaptic vesicles. Using this assay, high affinity saturable binding of recombinant βSpIIΣI proteins was observed with synaptic vesicles. Binding was only observed when the 25 amino acid synapsin I binding site was included on the recombinant peptides. Further, we demonstrate that antibodies directed against 15 amino acids of the synapsin I binding domain specifically blocked synaptic transmission in cultured hippocampal neurons. Thus, the synapsin I attachment site on βSpIIΣI spectrin comprises a ∼25 amino acid segment of the molecule and interaction of these two proteins is an essential step for the process of neurotransmission.