...
首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Solution structure of the ESCRT-I complex by small- angle X-ray scattering EPR and FRET spectroscopy
【24h】

Solution structure of the ESCRT-I complex by small- angle X-ray scattering EPR and FRET spectroscopy

机译:小角度X射线散射EPR和FRET光谱分析ESCRT-I配合物的溶液结构

获取原文
获取原文并翻译 | 示例
           

摘要

ESCRT-I is required for the sorting of integral membrane proteins to the lysosome, or vacuole in yeast, for cytokinesis in animal cells, and for the budding of HIV-1 from human macrophages and T lymphocytes. ESCRT-I is a heterotetramer. of Vps23, Vps28, Vps37, and Mvb12. The crystal structures of the core complex and the ubiquitin E2 variant and Vps28 C-terminal domains have been determined, but internal flexibility has prevented crystallization of intact ESCRT-I. Here we have characterized the structure of ESCRT-I in solution by simultaneous structural refinement against small-angle X-ray scattering and double electron-electron resonance spectroscopy of spin-labeled complexes. An ensemble of at least six structures, comprising an equally populated mixture of closed and open conformations, was necessary to fit all of the data. This structural ensemble was cross-validated against single-molecule FRET spectroscopy, which suggested the presence of a continuum of open states. ESCRT-I in solution thus appears to consist of an approximately 50% population of one or a few related closed conformations, with the other 50% populating a continuum of open conformations. These conformations provide reference points for the structural pathway by which ESCRT-I induces membrane buds.
机译:ESCRT-1是将完整的膜蛋白分选到溶酶体或酵母中的液泡,在动物细胞中进行胞质分裂以及从人巨噬细胞和T淋巴细胞中萌芽HIV-1所必需的。 ESCRT-1是异四聚体。 Vps23,Vps28,Vps37和Mvb12的数量。已经确定了核心复合物和遍在蛋白E2变体以及Vps28 C末端结构域的晶体结构,但是内部柔韧性阻止了完整ESCRT-1的结晶。在这里,我们通过针对小角度X射线散射的同时结构优化和自旋标记复合物的双电子-电子共振光谱,对溶液中ESCRT-I的结构进行了表征。为了拟合所有数据,必须包含至少六个结构的集合,包括闭合和开放构象的均等填充的混合物。该结构集成已针对单分子FRET光谱进行了交叉验证,这表明存在连续的开放状态。因此,溶液中的ESCRT-1似乎由大约50%的一种或几种相关的闭合构象组成,其他50%则构成了连续的开放构象。这些构象为ESCRT-1诱导膜芽的结构途径提供参考点。

著录项

  • 来源
  • 作者单位

    Laboratory of Molecular Biology and National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

    Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

    Department of Chemistry, Biophysics Program and Center for Membrane Biology, University of Virginia,Charlottesville, VA 22904-4319;

    Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

    Faculty of Mathematics and Physics, Institute of Physics, Charles University, 12116 Prague, Czech Republic;

    Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

    Department of Chemistry, Biophysics Program and Center for Membrane Biology, University of Virginia,Charlottesville, VA 22904-4319;

    Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

    Laboratory of Molecular Biology and National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    hybrid methods; protein sorting; vesicle budding; multiprotein assemblies; ensemble refinement;

    机译:混合方法蛋白质分选;囊泡出芽;多蛋白组装;合奏细化;

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