Wiser Ofer, Cohen Roy, Atlas Daphne
Department of Biological Chemistry Institute of Life Sciences, Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3968-73. doi: 10.1073/pnas.052017299. Epub 2002 Mar 12.
Alteration of the kinetic properties of voltage-gated Ca(2+) channels, Ca(v)1.2 (Lc-type), Ca(v)2.2 (N type), and Ca(v)2.3 (R type), by syntaxin 1A (Syn1A) and synaptotagmin could modulate exocytosis. We tested how switching divalent charge carriers from Ca(2+) to Sr(2+) and Ba(2+) affected Syn1A and synaptotagmin modulation of Ca(2+)-channel activation. Syn1A accelerated Ca(v)1.2 activation if Ca(2+) was the charge carrier; and by substituting for Ba(2+), Syn1A slowed Ca(v)1.2 activation. Syn1A also significantly accelerated Ca(v)2.3 activation in Ca(2+) and marginally in Ba(2+). Synaptotagmin, on the other hand, increased the rate of activation of Ca(v)2.3 and Ca(v)2.2 in all permeating ions tested. The Syn1A-channel interaction, unlike the synaptotagmin-channel interaction, proved significantly more sensitive to the type of permeating ion. It is well established that exocytosis is affected by switching the charge carriers. Based on the present results, we suggest that the channel-Syn1A interaction could respond to the conformational changes induced within the channel during membrane depolarization and divalent ion binding. These changes could partially account for the charge specificity of synaptic transmission as well as for the fast signaling between the Ca(2+) source and the fusion apparatus of channel-associated-vesicles (CAV). Furthermore, propagation of conformational changes induced by the divalent ions appear to affect the concerted interaction of the channel with the fusion/docking machinery upstream to free Ca(2+) buildup and/or binding to a cytosolic Ca(2+) sensor. These results raise the intriguing possibility that the channel is the Ca(2+) sensor in the process of fast neurotransmitter release.
Syntaxin 1A(Syn1A)和突触结合蛋白对电压门控Ca(2+)通道Ca(v)1.2(L型)、Ca(v)2.2(N型)和Ca(v)2.3(R型)动力学特性的改变可调节胞吐作用。我们测试了将二价电荷载体从Ca(2+)切换为Sr(2+)和Ba(2+)如何影响Syn1A和突触结合蛋白对Ca(2+)通道激活的调节。如果Ca(2+)是电荷载体,Syn1A会加速Ca(v)1.2的激活;而通过用Ba(2+)替代,Syn1A会减慢Ca(v)1.2的激活。Syn1A在Ca(2+)中也显著加速Ca(v)2.3的激活,在Ba(2+)中略有加速。另一方面,突触结合蛋白在所有测试的通透离子中都增加了Ca(v)2.3和Ca(v)2.2的激活速率。与突触结合蛋白-通道相互作用不同,Syn1A-通道相互作用对通透离子类型的敏感性明显更高。众所周知,胞吐作用会受到电荷载体切换的影响。基于目前的结果,我们认为通道-Syn1A相互作用可能对膜去极化和二价离子结合过程中通道内诱导的构象变化做出反应。这些变化可能部分解释了突触传递的电荷特异性以及Ca(2+)源与通道相关囊泡(CAV)融合装置之间的快速信号传导。此外,由二价离子诱导的构象变化的传播似乎会影响通道与融合/对接机制在游离Ca(2+)积累和/或与胞质Ca(2+)传感器结合上游的协同相互作用。这些结果提出了一个有趣的可能性,即通道是快速神经递质释放过程中的Ca(2+)传感器。