OXIDATION-INDUCED INTRAMOLECULAR DISULFIDE BOND INACTIVATES MKK6

氧化诱导的分子内二硫键使 MKK6 失活

• 批准号: 8365917
• 负责人: WUA ZHENGUO
• 金额: $ 1.28万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365917
• 关键词: Binding; Biochemical; Biology; Cellular Assay; Cysteine; Disulfides; Family; Funding; Fungal Genome; Grant; MAP Kinase Kinase 6; MAPK14 gene; Mitogen-Activated Protein Kinase Kinases; National Center for Research Resources; Oxidation-Reduction; Phosphotransferases; Principal Investigator; Reducing Agents; Research; Research Infrastructure; Resources; Source; United States National Institutes of Health; Work; base; cost; disulfide bond; member; oxidation; sensor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Mitogen-activated protein kinase kinase 6 (MKK6) is a member of the mitogen-activated protein kinase (MAPK) kinase (MAP2K) subfamily that specifically phosphorylates and activates the p38 MAPKs. Based on both biochemical and cellular assays, we found that MKK6 was extremely sensitive to oxidation: It was inactivated by oxidation and its kinase activity was fully restored upon treatment with a reducing agent. Detailed mechanistic studies showed that cysteines 109 and 196, two of the six cysteines in MKK6, formed an intramolecular disulfide bond upon oxidation that inactivated MKK6 by inhibiting its ATP binding. This mechanism is distinct from that seen in other redox-sensitive kinases. The two cysteines involved in intramolecular disulfide formation are conserved in all seven members of the MAP2K family. Consistently, we confirmed that other MAP2Ks were also sensitive to oxidation. Our work reveals that MKK6 and other MAP2Ks are a distinct class of cellular redox sensors.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 子项目的主要研究者可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 促分裂原活化蛋白激酶激酶6（MKK 6）是促分裂原活化蛋白激酶（MAPK）激酶（MAP 2K）亚家族的成员，其特异性磷酸化并活化p38 MAPK。基于生物化学和细胞测定，我们发现MKK 6对氧化极其敏感：它被氧化灭活，并且在用还原剂处理后其激酶活性完全恢复。详细的机制研究表明，半胱氨酸109和196（MKK 6中六个半胱氨酸中的两个）在氧化后形成分子内二硫键，通过抑制ATP结合使MKK 6失活。这种机制与其他氧化还原敏感性激酶中所见的机制不同。参与分子内二硫键形成的两个半胱氨酸在MAP 2K家族的所有七个成员中是保守的。因此，我们证实了其他MAP 2Ks也对氧化敏感。我们的工作表明，MKK 6和其他MAP 2Ks是一类独特的细胞氧化还原传感器。