Rapid assay of binding of tumor-promoting phorbol esters to protein kinase C1.
Rapid assay of binding of tumor-promoting phorbol esters to protein kinase C1.
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快速测定促进肿瘤的佛波酯与蛋白激酶 C1 的结合。
DOI：
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发表时间：
1986
期刊：
影响因子：
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通讯作者：
Y. Nishizuka
中科院分区：
文献类型：
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作者：
Y. Tanaka;R. Miyake;U. Kikkawa;Y. Nishizuka
蛋白激酶C被普遍认为是促肿瘤佛波酯的受体蛋白。[3 H]佛波醇-12，13-二丁酸酯与蛋白激酶C的结合可通过快速过滤程序测定，该程序使用已用阳离子聚合物聚乙烯亚胺处理的玻璃纤维过滤器。佛波醇酯仅在磷脂酰丝氨酸和钙存在下特异性结合蛋白激酶。非特异性结合最多小于总结合的10%。结合相对于蛋白激酶C的浓度是线性的，依赖于佛波酯和磷脂酰丝氨酸的浓度以饱和的方式，并被二酰基甘油（蛋白激酶的内源性激活剂）抑制。
Protein kinase C is generally accepted to be a receptor protein of tumor-promoting phorbol esters. The binding of [3H]phorbol-12,13-dibutyrate to protein kinase C can be assayed by a rapid filtration procedure using a glass-fiber filter that has been treated with a cationic polymer, polyethylenimine. The phorbol ester specifically binds to the protein kinase only in the presence of phosphatidylserine and calcium. Non-specific binding is less than 10%, at most, of the total binding. The binding is linear with respect to the concentration of protein kinase C, is dependent on the concentrations of phorbol ester and phosphatidylserine in a saturative manner, and is inhibited by diacylglycerol (an endogenous activator of the protein kinase).