Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry.

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry.
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DOI:
10.3791/57966
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发表时间:
2018-10-15
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Politis A
Politis A
中科院分区:
其他
文献类型:
--
作者:
Ahdash Z;Lau AM;Martens C;Politis A

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蛋白质是一类重要的生物大分子,在细胞功能中起着许多关键作用,包括基因表达、催化代谢反应、DNA修复和复制。因此,对这些过程的详细了解提供了关于细胞功能的关键信息。综合结构质谱方法提供蛋白质复合物组装、复合物连通性、亚基化学计量学、蛋白质寡聚化和配体结合的结构和动态信息。综合结构质谱的最新进展已经允许表征具有挑战性的生物系统,包括大的DNA结合蛋白和膜蛋白。该方案描述了如何整合不同的质谱数据,如原生质谱和离子迁移-质谱(IM-MS)与分子动力学模拟,以深入了解解旋酶-核酸酶DNA修复蛋白复合物。由此产生的方法为参与重要生物过程的配体与其他蛋白质复合物结合的详细研究提供了框架。
Proteins are an important class of biological macromolecules that play many key roles in cellular functions including gene expression, catalyzing metabolic reactions, DNA repair and replication. Therefore, a detailed understanding of these processes provides critical information on how cells function. Integrative structural MS methods offer structural and dynamical information on protein complex assembly, complex connectivity, subunit stoichiometry, protein oligomerization and ligand binding. Recent advances in integrative structural MS have allowed for the characterization of challenging biological systems including large DNA binding proteins and membrane proteins. This protocol describes how to integrate diverse MS data such as native MS and ion mobility-mass spectrometry (IM-MS) with molecular dynamics simulations to gain insights into a helicase-nuclease DNA repair protein complex. The resulting approach provides a framework for detailed studies of ligand binding to other protein complexes involved in important biological processes.
完整膜蛋白复合物的质谱法。
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