...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Crystallographic Snapshots of Tom20–Mitochondrial Presequence Interactions with Disulfide-Stabilized Peptides
【24h】

Crystallographic Snapshots of Tom20–Mitochondrial Presequence Interactions with Disulfide-Stabilized Peptides

机译：Tom20 –线粒体序列相互作用与二硫键稳定的肽的晶体快照。

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

Most mitochondrial proteins are synthesized in thencytosol and imported into mitochondria. The Tom20 protein,nresiding on the mitochondrial surface, recognizes the N-terminalnpresequences of precursor proteins. We previously determined thencrystal structures of the Tom20u0001presequence complex. The success-nful crystallization involved tethering the presequence to Tom20nthrough an intermolecular disulfide bond with an optimized linker.nIn this work, we assessed the tethering method. The intermolecularndisulfide bond was cleaved in crystal with a reducing agent. The posen(i.e., conformation and position) of the presequence was identical to the previously determined pose. In another experiment, anlonger linker than the optimized length was used for the tethering. The perturbation of the tether changed the pose slightly, but theninteraction mode was preserved. These results argue against the forced interaction of the presequence by its covalent attachment tonTom20. Second, as an alternative method referred to as “molecular stiffening”, we introduced a disulfide bond within thenpresequence peptide to restrict the freedomof the peptide in the unbound states.One presequence analogue exhibited over 100-foldnhigher affinity than its linear counterpart and generated cocrystals with Tom20.One of the two crystallographic snapshots revealed anknown pose previously determined by the tethering method, and the other snapshot depicted a new pose. These results confirmednand extended the dynamic, multiple bound state model of the Tom20u0001presequence interactions and also demonstrated the validitynof the molecular tethering and stiffening techniques in studies of transient proteinu0001peptide interactions.
机译：大多数线粒体蛋白在胞浆中合成，然后导入线粒体。位于线粒体表面的Tom20蛋白可识别前体蛋白的N端n序列。我们先前确定了Tom20u0001presequence配合物的晶体结构。成功的结晶涉及通过分子间二硫键和优化的连接子将序列连接到Tom20n。在这项工作中，我们评估了连接方法。分子间二硫键被还原剂裂解成晶体。序列的姿势（即构象和位置）与先前确定的姿势相同。在另一个实验中，将比最佳长度更长的接头用于束缚。系绳的扰动稍微改变了姿势，但随后保留了交互模式。这些结果证明了共价键tonTom20迫使先序列相互作用。其次，作为一种称为“分子增稠”的替代方法，我们在先序列肽中引入了二硫键以限制肽在未结合状态下的自由。一个先序列类似物的亲和力比其线性对应物高100倍，并生成共晶体。两个晶体快照中的一个快照揭示了一个以前通过束缚方法确定的已知姿势，另一个快照描述了一个新的姿势。这些结果证实并扩展了Tom20u0001前序列相互作用的动态多束缚状态模型，并证明了分子束缚和强化技术在瞬时蛋白u0001肽相互作用研究中的有效性。

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