PROTEIN-KINASE-C DOMAINS INVOLVED IN INTERACTIONS WITH OTHER PROTEINS
PROTEIN-KINASE-C DOMAINS INVOLVED IN INTERACTIONS WITH OTHER PROTEINS
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DOI：
10.1021/bi00171a024
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发表时间：
1994-02-08
期刊：
影响因子：
2.9
通讯作者：
JAKEN, S
中科院分区：
文献类型：
--
作者：
LIAO, L;HYATT, SL;JAKEN, S
我们已经使用了印迹覆盖试验来检测蛋白激酶C（PKC）与其他蛋白质的相互作用。在许多情况下，PKC结合蛋白也是PKC底物[Chapline等人（1993）J. biol. Chem. 268，6858]。目前研究的目的是表征参与与其他蛋白质相互作用的PKC结构域，PKC的α、β和β亚型与成纤维细胞提取物中相同的结合蛋白相互作用。这些结果表明，恒定的，而不是同工酶特异性（可变）区域是研究的相互作用的主要决定因素。PKC的结合需要磷脂酰丝氨酸（PS），这表明PKC的PS结合调节结构域参与了相互作用。PKC假底物肽序列，这是包含在调节结构域中，也显示PS依赖性的促进PKC-蛋白质相互作用，N-末端截短突变体缺乏的假底物序列的制备。与野生型α-PKC相比，突变型α-PKC的结合减少，尽管一些结合仍然明显。这些结果表明，假底物序列有助于，但不是唯一的决定因素，PKC结合活性。
We have used a blot overlay assay to detect protein kinase C (PKC) interactions with other proteins. In many cases, the PKC binding proteins are also PKC substrates [Chapline et al. (1993) J. biol. Chem. 268, 6858]. The purpose of the current studies was to characterize the PKC domains involved in the interactions with other proteins, alpha, beta, and epsilon isoforms of PKC interact with the same binding proteins in fibroblast cell extracts. These results indicate that constant rather than isozyme-specific (variable) regions are the major determinants of the interactions studied. PKC binding required phosphatidylserine (PS), indicating that the PS binding regulatory domain of PKC is involved in the interactions. The PKC pseudosubstrate peptide sequence, which is contained within the regulatory domain, also showed PS-dependent in promoting PKC-protein interactions, an N-terminal truncation mutant lacking the pseudosubstrate sequence was prepared. Binding of the mutant alpha-PKC was diminished compared to wild-type alpha-PKC, although some binding was still apparent. These results that the pseudosubstrate sequence contributes to, but is not the sole determinant of, PKC binding activity.