Rational design of a selective antagonist of ε protein kinase C derived from the selective allosteric agonist, pseudo-RACK peptide
Rational design of a selective antagonist of ε protein kinase C derived from the selective allosteric agonist, pseudo-RACK peptide
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DOI：
10.1016/j.yjmcc.2007.01.007
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发表时间：
2007-04-01
影响因子：
5
通讯作者：
Mochly-Rosen, Daria
中科院分区：
文献类型：
--
作者：
Liron, Tamar;Chen, Leon E.;Mochly-Rosen, Daria
我们以前已经表明，结构域参与结合蛋白激酶C（PKC）同工酶，其各自的锚定蛋白（RACK）和短肽来自这些结构域的PKC同工酶选择性拮抗剂。我们还确定了PKC同工酶选择性激动剂，命名为psi RACK肽，来自每个PKC内的序列与其各自的RACK具有高度同源性。我们注意到，每个PKC同工酶内的所有psi RACK序列与其相应的RACK至少有一个非同源氨基酸差异，这构成了电荷变化。基于这些信息，我们在这里设计了一种新的方法来设计一种同工酶选择性PKC拮抗剂，来自psi RACK序列。我们集中于pSKPKC psi RACK肽，其中假pSKRACK序列（pSKRACK;柱APIGYD下的H（D）;对应于pSKPKC 85 -92）在第二个氨基酸中与同源RACK衍生序列（柱VALGYD下的N（N）;对应于pSKRACK 285 -292）的电荷不同。在这里，我们表明，通过仅一个氨基酸的取代（天冬氨酸到天冬酰胺）改变psi RACK肽的电荷导致在相同的细胞功能上具有相反活性的肽，并且用丙氨酸取代天冬氨酸导致无活性肽。这些数据支持了我们关于伪RACK肽激活心脏细胞中PKC的机制的假设，并表明这种方法适用于具有诱导蛋白质-蛋白质相互作用的其他信号蛋白。(c)2007年爱思唯尔公司All rights reserved.
We have previously shown that domains involved in binding of protein kinase C (PKC) isozymes to their respective anchoring proteins (RACKs) and short peptides derived from these domains are PKC isozyme-selective antagonists. We also identified PKC isozyme-selective agonists, named psi RACK peptides, derived from a sequence within each PKC with high homology to its respective RACK. We noted that all the psi RACK sequences within each PKC isozyme have at least one non-homologous amino acid difference from their corresponding RACK that constitutes a charge change. Based on this information, we have devised here a new approach to design an isozyme-selective PKC antagonist, derived from the psi RACK sequence. We focused on epsilon PKC psi RACK peptide, where the pseudo-epsilon RACK sequence (psi epsilon RACK; H (D) under bar APIGYD; corresponding to epsilon PKC85-92) is different in charge from the homologous RACK-derived sequence (N (N) under bar VALGYD; corresponding to epsilon RACK285-292) in the second amino acid. Here we show that changing the charge of the psi epsilon RACK peptide through a substitution of only one amino acid (aspartate to asparagine) resulted in a peptide with an opposite activity on the same cell function and a substitution for aspartate with an alanine resulted in an inactive peptide. These data support our hypothesis regarding the mechanism by which pseudo-RACK peptide activates PKC in heart cells and suggest that this approach is applicable to other signaling proteins with inducible protein-protein interactions. (c) 2007 Elsevier Inc. All rights reserved.