LOW-AFFINITY FINDING OF PHORBOL ESTERS TO PROTEIN-KINASE-C AND ITS RECOMBINANT CYSTEINE-RICH REGION IN THE ABSENCE OF PHOSPHOLIPIDS

被引:71
作者
KAZANIETZ, MG
BARCHI, JJ
OMICHINSKI, JG
BLUMBERG, PM
机构
[1] NCI,MED CHEM LAB,BETHESDA,MD 20892
[2] NIDDK,CHEM PHYS LAB,BETHESDA,MD 20892
[3] NCI,CELLULAR CARCINOGENESIS & TUMOR PROMOT LAB,MOLEC MECHANISMS TUMOR PROMOT SECT,BETHESDA,MD 20892
关键词
D O I
10.1074/jbc.270.24.14679
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Binding of phorbol esters to protein kinase C (PKC) has been regarded as dependent on phospholipids, with phosphatidylserine being the most effective for reconstituting binding. By using a purified single cysteine-rich region from PKC delta expressed in Escherichia coli we were able to demonstrate that specific binding of [H-3]phorbol 12,13-dibutyrate to the receptor still takes place in the absence of the phospholipid cofactor. However, [H-3]phorbol 12,13-dibutyrate bound to the cysteine rich region with 80-fold lower affinity in the absence than in the presence of 100 mu g/ml phosphatidlylserine. Similar results were observed with the intact recombinant PRC delta isolated from insect cells. When different phorbol derivatives were examined, distinct structure-activity relations for the cysteine-rich region were found in the presence and absence of phospholipid. Our results have potential implications for PKC translocation, for inhibitor design, and for PKC structural determination.
引用
收藏
页码:14679 / 14684
页数:6
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