Identification of the critical features of a small peptide inhibitor of JNK activity
Identification of the critical features of a small peptide inhibitor of JNK activity
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DOI：
10.1074/jbc.m107565200
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发表时间：
2002-03-29
影响因子：
4.8
通讯作者：
Bogoyevitch, MA
中科院分区：
文献类型：
--
作者：
Barr, RK;Kendrick, TS;Bogoyevitch, MA
c-Jun N-末端激酶（JNK）是丝裂原活化蛋白激酶（NLAPK）的一个亚家族。尽管通过特异性抑制剂的表征促进了对其他MAPK功能的评估，但没有JNK定向抑制剂可商购获得。基于JNK支架蛋白JNK相互作用蛋白-1（JIP-1）的JNK结合结构域（JBD）的氨基酸143-163，我们已经鉴定了活化的JNK的21个氨基酸的肽抑制剂。这种肽，I-JIP（基于JIP-1的JNK抑制剂），在体外抑制JNK对重组c-Jun、Elk和ATF 2的活性高达90%。表面等离子体共振分析表明，截短的I-JIP（TI-JIP），即I-JIP的C-末端11个氨基酸，直接与重组JNK相互作用，但不与其底物相互作用。在截短的I-JIP内扫描丙氨酸置换鉴定了4个残基（Arg-156、Pro-157、Leu-160或Leu-162）作为独立的抑制关键残基。来自JIP-2和JIP-3的JBD肽序列共享这些关键残基，因此是有效的JNK抑制剂。相比之下，基于ATF 2和c-Jun的JBD的肽通过抑制JNK活性，
The c-Jun N-terminal kinases (JNKs) are a subfamily of the mitogen-activated protein kinases (NLAPKs). Although progress in evaluating the functions of other MAPKs has been facilitated by the characterization of specific inhibitors, no JNK-directed inhibitor is commercially available. We have identified a 21-amino acid peptide inhibitor of activated JNKs, based on amino acids 143-163 of the JNK-binding domain (JBD) of the JNK scaffolding protein, JNK-interacting protein-1 (JIP-1). This peptide, I-JIP (Inhibitor of JNK-based on JIP-1), inhibited JNK activity in vitro toward recombinant c-Jun, Elk, and ATF2 up to 90%. A truncated I-JIP (TI-JIP), the C-terminal 11 amino acids of I-JIP, directly interacted with recombinant JNKs but not its substrates as shown by surface plasmon resonance analysis. Scanning alanine replacement within truncated I-JIP identified 4 residues (Arg-156, Pro-157, Leu-160, or Leu-162) as independently critical for inhibition. JBD peptide sequences from JIP-2 and JIP-3 shared these critical residues and accordingly were effective JNK inhibitors. In contrast, peptides based on the JBDs of ATF2 and c-Jun inhibited JNK activity by