...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Energetics and specificity of interactions within Ub center dot Uev center dot Ubc13 human ubiquitin conjugation complexes
【24h】

Energetics and specificity of interactions within Ub center dot Uev center dot Ubc13 human ubiquitin conjugation complexes

机译：Ub中心点Uev中心点Ubc13人泛素结合复合物内相互作用的能量学和特异性

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Lys(63)-linked polyubiquitin (poly-Ub) chains appear to play a nondegradative signaling and/or recruitment role in a variety of key eukaryotic cellular processes, including NF-kappaB signal transduction and DNA repair. A protein heterodimer composed of a catalytically active ubiquitin-conjugating enzyme (Ubc13) and its homologue (Mms2 or Uev1a) forms a catalytic scaffold upon which a noncovalently associated acceptor Ub and thiolester-linked donor Ub are oriented such that LyS(63)-linked poly-Ub chain synthesis is facilitated. In this study, we have used H-1-N-15 nuclear magnetic resonance spectroscopy, in combination with isothermal titration calorimetry, to determine the thermodynamics and kinetics of the interactions between various components of the LyS63-linked poly-Ub conjugation machinery. Mms2 and Uev1a interact in vitro with acceptor Ub to form 1/1 complexes with macroscopic dissociation constants of 98 +/- 15 and 213 +/- 14 muM, respectively, and appear to bind Ub in a similar fashion. Interestingly, the Mms2(.)Ubc13 heterodimer associates with acceptor Ub in a 1/1 complex and binds with a dissociation constant of 28 +/- 6 muM, significantly stronger than the binding of Mms2 alone. Furthermore, a dissociation constant of 49 +/- 7 nM was determined for the interaction between Mms2 and Ubc13 using isothermal titration calorimetry. In connection with previous structural studies for this system, the thermodynamics and kinetics of acceptor Ub binding to the Mms2(.)Ubc13 heterodimer described in detail in this study will allow for a more thorough rationalization of the mechanism of formation of LyS(63)-linked poly-Ub chains. [References: 38]
机译：与Lys（63）连接的聚泛素（poly-Ub）链似乎在各种关键的真核细胞过程（包括NF-κB信号转导和DNA修复）中发挥非降解的信号传导和/或募集作用。由具有催化活性的泛素结合酶（Ubc13）和其同系物（Mms2或Uev1a）组成的蛋白质异二聚体形成催化支架，非共价结合的受体Ub和与硫酯连接的供体Ub定向在该支架上，从而使LyS（63）连接聚-Ub链的合成变得容易。在这项研究中，我们将H-1-N-15核磁共振波谱与等温滴定量热法结合使用，确定了与LyS63连接的多Ub偶联机械中各个组分之间相互作用的热力学和动力学。 Mms2和Uev1a在体外与受体Ub相互作用形成1/1复合物，其宏观解离常数分别为98 +/- 15和213 +/- 14μM，并且似乎以相似的方式结合Ub。有趣的是，Mms2（。）Ubc13异二聚体与受体Ub以1/1复合物缔合，并以28 +/- 6μM的解离常数结合，明显强于单独的Mms2。此外，使用等温滴定量热法测定了Mms2和Ubc13之间的相互作用的解离常数为49 +/- 7 nM。与该系统先前的结构研究相关，本研究中详细描述的受体Ub与Mms2（。）Ubc13异二聚体结合的热力学和动力学将使LyS（63）-的形成机理更彻底合理化。链接的多Ub链。 [参考：38]

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号