Regulated Binding of the Protein Kinase C Substrate GAP-43 to the V0/C2 Region of Protein Kinase C-δ*
Regulated Binding of the Protein Kinase C Substrate GAP-43 to the V0/C2 Region of Protein Kinase C-δ*
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蛋白激酶 C 底物 GAP-43 与蛋白激酶 C-δ* V0/C2 区域的调节结合
DOI：
10.1074/jbc.272.19.12747
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
P. Parker
中科院分区：
文献类型：
--
作者：
L. Dekker;P. Parker
研究了蛋白激酶C-δ与其神经底物GAP-43之间的相互作用。从COS细胞中分离出两种形式的蛋白激酶C-δ，其特征在于凝胶迁移率、GAP-43结合以及特异性GAP-43和组蛋白激酶活性的差异。一种缓慢迁移、低比活性形式的蛋白激酶C-δ直接与固定化GAP-43结合。在EGTA的存在下，结合被废除，这表明相互作用的Ca 2+依赖性。蛋白激酶C-δ的游离催化结构域不与GAP-43结合，表明在调节结构域中存在结合位点。产生谷胱甘肽S-转移酶-蛋白激酶C-δ调节结构域融合蛋白，并测试其与GAP-43的结合。V0/C2样氨基末端结构域被定义为差距-43结合位点。GAP-43与该区域的结合被EGTA抑制，并在10−7和10−6 m之间的Ca 2+水平下调节。通过在人胚肾细胞中共表达蛋白激酶C-δ和GAP-43，然后通过免疫沉淀，在完整细胞中研究了这两种蛋白之间的相互作用。复合物的形成仅发生在用Ca 2+离子载体离子霉素处理细胞后，表明细胞内Ca 2+的升高是体内相互作用所需的。它的结论是，蛋白激酶C-δ与GAP-43通过V0/C2-样结构域，催化位点外，这种相互作用是由细胞内Ca 2+调节。
The interaction between protein kinase C-δ and its neuronal substrate, GAP-43, was studied. Two forms of protein kinase C-δ were isolated from COS cells and characterized by differences in gel mobility, GAP-43 binding, and specific GAP-43 and histone kinase activities. A slow migrating, low specific activity form of protein kinase C-δ bound directly to immobilized GAP-43. Binding was abolished in the presence of EGTA, suggesting Ca2+ dependence of the interaction. The free catalytic domain of protein kinase C-δ did not bind GAP-43, suggesting the existence of a binding site in the regulatory domain. Glutathione S-transferase-protein kinase C-δ regulatory domain fusion proteins were generated and tested for binding to GAP-43. The V0/C2-like amino-terminal domain was defined as the GAP-43-binding site. GAP-43 binding to this region is inhibited by EGTA and regulated at Ca2+ levels between 10−7 and 10−6 m. The interaction between protein kinase C-δ and GAP-43 was studied in intact cells by coexpression of the two proteins in human embryonic kidney cells followed by immunoprecipitation. Complex formation occurred only after treatment of the cells with the Ca2+ ionophore ionomycin, indicating that elevation of intracellular Ca2+ is required for interaction in vivo. It is concluded that protein kinase C-δ interacts with GAP-43 through the V0/C2-like domain, outside the catalytic site, and that this interaction is modulated by intracellular Ca2+.
神经元特异性蛋白 F1/GAP-43 对蛋白激酶 C 的 β 亚型具有底物特异性。
DOI：
10.1016/0006-291x(90)90818-8
发表时间：
1990
影响因子：
3.1
作者：
Sheu,FS;Marais,RM;Parker,PJ;Bazan,NG;Routtenberg,A
通讯作者：
Routtenberg,A
影响因子：
56.9
作者：
MOCHLYROSEN, D
通讯作者：
MOCHLYROSEN, D
影响因子：
2.9
作者：
LIAO, L;HYATT, SL;JAKEN, S
通讯作者：
JAKEN, S