Nelson T J, Cavallaro S, Yi C L, McPhie D, Schreurs B G, Gusev P A, Favit A, Zohar O, Kim J, Beushausen S, Ascoli G, Olds J, Neve R, Alkon D L
Laboratory of Adaptive Systems, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA.
Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13808-13. doi: 10.1073/pnas.93.24.13808.
A previously uncharacterized 22-kDa Ca(2+)-binding protein that also binds guanosine nucleotides was characterized, cloned, and analyzed by electrophysiological techniques. The cloned protein, calexcitin, contains two EF-hands and also has homology with GTP-binding proteins in the ADP ribosylation factor family. In addition to binding two molecules of Ca2+, calexcitin bound GTP and possessed GTPase activity. Calexictin is also a high affinity substrate for protein kinase C. Application of calexcitin to the inner surface of inside-out patches of human fibroblast membranes, in the presence of Ca2+ and the absence of endogenous Ca2+/calmodulin kinase type II or protein kinase C activity, reduced the mean open time and mean open probability of 115 +/- 6 pS K+ channels. Calexcitin thus appears to directly regulate K+ channels. When microinjected into molluscan neurons or rabbit cerebellar Purkinje cell dendrites, calexcitin was highly effective in enhancing membrane excitability. Because calexcitin translocates to the cell membrane after phosphorylation, calexcitin could serve as a Ca(2+)-activated signaling molecule that increases cellular excitability, which would in turn increase Ca2+ influx through the membrane. This is also the first known instance of a GTP-binding protein that binds Ca2+.
一种先前未被鉴定的22 kDa钙结合蛋白,该蛋白也能结合鸟苷核苷酸,通过电生理技术对其进行了表征、克隆和分析。克隆得到的蛋白——钙激活蛋白,含有两个EF手结构,并且与ADP核糖基化因子家族中的GTP结合蛋白具有同源性。除了能结合两个钙离子分子外,钙激活蛋白还能结合GTP并具有GTP酶活性。钙激活蛋白也是蛋白激酶C的高亲和力底物。在存在钙离子且不存在内源性II型钙/钙调蛋白激酶或蛋白激酶C活性的情况下,将钙激活蛋白应用于人成纤维细胞膜内翻片的内表面,可降低115±6 pS钾通道的平均开放时间和平均开放概率。因此,钙激活蛋白似乎能直接调节钾通道。当将钙激活蛋白显微注射到软体动物神经元或兔小脑浦肯野细胞树突中时,它能非常有效地增强膜兴奋性。由于钙激活蛋白在磷酸化后会转位到细胞膜,它可以作为一种钙激活信号分子来增加细胞兴奋性,进而增加钙离子通过膜的内流。这也是已知的第一个能结合钙离子的GTP结合蛋白的例子。