N-type voltage-sensitive calcium channel interacts with syntaxin, synaptotagmin and SNAP-25 in a multiprotein complex.

Tobi D; Wiser O; Trus MAtlas D

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N-type voltage-sensitive calcium channel interacts with syntaxin, synaptotagmin and SNAP-25 in a multiprotein complex.

机译：N-type voltage-sensitive calcium channel interacts with syntaxin, synaptotagmin and SNAP-25 in a multiprotein complex.

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Expression of the N-type voltage sensitive calcium channel in Xenopus oocytes along with syntaxin and p65 showed that the syntaxin-modified N-type channel properties, were fully reversed by p65. The inward current was restored to a significantly higher amplitude when all three proteins were present, suggesting that the channel interacts with syntaxin, p65 and SNAP-25 in a quaternary complex. Further support to a multicomplex formation between the channel and the synaptic proteins was drawn from the steady-state voltage inactivation profiles. A physical interaction of the N-type calcium channel with the vesicular protein synaptotagmin (p65) was demonstrated biochemically, using recombinant fusion proteins. The interaction is confined to a cytosolic channel domain that separates segments II and III amino acids 710-1090 of the N-type channel (N-Loop710-1090). In vitro binding of recombinant N-Loop710-1090 to p65 (amino acids 96-421) involves the two C2 domains of p65, C2A domain amino acids 96-265; p65(1-3) and C2B domain amino acids 248-421; p65(3-5). While the binding of C2A and C2B domains was calcium independent, C2B domain binding to the N-Loop was inositol-hexaphosphate (IP6)-sensitive. The N-Loop710-1090 binding to p65 was competed by syntaxin and SNAP-25, which are synaptic plasma membrane proteins. These combined functional and biochemical approaches provide evidence for a complex formation between the N-type channel and the exocytotic machinery which by generating fusion-competent vesicles may function to regulate the process of synaptic secretion.

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《Receptors and channels》 |1998年第2期|89-98|共10页
作者
Tobi D; Wiser O; Trus MAtlas D;
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Department of Biological Chemistry, Hebrew University of Jerusalem, Israel.;

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正文语种英语
中图分类接收设备、无线电收音机;
关键词
Calcium Channels; Ion Channel Gating; Membrane Glycoproteins; Membrane Proteins; 钙通道; 离子通道闸门; 膜糖蛋白类; 膜蛋白质类; 神经组织蛋白质类;

N-type voltage-sensitive calcium channel interacts with syntaxin, synaptotagmin and SNAP-25 in a multiprotein complex.

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