Substrate recognition domains within extracellular signal-regulated kinase mediate binding and catalytic activation of mitogen-activated protein kinase phosphatase-3
Substrate recognition domains within extracellular signal-regulated kinase mediate binding and catalytic activation of mitogen-activated protein kinase phosphatase-3
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DOI：
10.1074/jbc.m001515200
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发表时间：
2000-08-11
影响因子：
4.8
通讯作者：
Arkinstall, S
中科院分区：
文献类型：
--
作者：
Nichols, A;Camps, M;Arkinstall, S
丝裂原活化蛋白激酶磷酸酶-3（MKP-3）是一种双特异性磷酸酶，可使细胞外信号调节激酶（ERK）MAP激酶失活。这反映了ERR和MKP-8氨基末端之间的紧密和特异性结合，随后磷酸酶活化和结合的MAP激酶的去磷酸化。我们已经使用了一系列p38/ERK嵌合分子来鉴定ERR内结合和催化活化MKP-3所必需的结构域。这些研究表明，ERK激酶亚结构域V-XI对于MKP-3的结合和催化活化是必要的和充分的。这些结构域构成ERK的主要COOH末端结构叶，具有这些区域的p38/ERK嵌合体显示对MKP-3失活的敏感性增加。这些数据还揭示了与MKP-3相互作用的ERR结构域与已知赋予ERK RAP激酶底物特异性的ERR结构域之间的重叠。与此相一致，我们表明，肽代表对接点内的目标基板Elk-1和p90（rsk）抑制ERR依赖性激活MKP-3。此外，在MKP-3 NH 2末端内bar酶（i）下bar相互作用（m）下bar互作（m）下bar酶（k）突变后，ERR依赖性磷酸酶活化消失，表明ERK COOH末端结构叶的关键接触位点包括位于DSP基因家族成员之间保守的Cdc 25同源结构域（CH 2）之间的残基。
Mitogen-activated protein (MAP) kinase phosphatase-3 (MKP-3) is a dual specificity phosphatase that inactivates extracellular signal-regulated kinase (ERK) MAP kinases. This reflects tight and specific binding between ERR and the MKP-8 amino terminus with consequent phosphatase activation and dephosphorylation of the bound MAP kinase. We have used a series of p38/ERK chimeric molecules to identify domains within ERR necessary for binding and catalytic activation of MKP-3. These studies demonstrate that ERK kinase subdomains V-XI are necessary and sufficient for binding and catalytic activation of MKP-3. These domains constitute the major COOH-terminal structural lobe of ERK, p38/ERK chimeras possessing these regions display increased sensitivity to inactivation by MKP-3. These data also reveal an overlap between ERR domains interacting with MKP-3 and those known to confer substrate specificity on the ERK RAP kinase. Consistent with this, we show that peptides representing docking sites within the target substrates Elk-1 and p90(rsk) inhibit ERR-dependent activation of MKP-3. In addition, abolition of ERR-dependent phosphatase activation following mutation of a putative (k) under bar inase (i) under bar nteraction (m) under bar otif (KIM) within the MKP-3 NH2 terminus suggests that key sites of contact for the ERK COOH-terminal structural lobe include residues localized between the Cdc25 homology domains (CH2) found conserved between members of the DSP gene family.