Ahdash Zainab, Lau Andy M, Martens Chloe, Politis Argyris
Department of Chemistry, King's College London.
Department of Chemistry, King's College London;
J Vis Exp. 2018 Oct 15(140):57966. doi: 10.3791/57966.
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.
蛋白质是一类重要的生物大分子，在细胞功能中发挥着许多关键作用，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为细胞如何发挥功能提供了关键信息。整合结构质谱方法可提供有关蛋白质复合物组装、复合物连接性、亚基化学计量、蛋白质寡聚化和配体结合的结构和动力学信息。整合结构质谱的最新进展使得对具有挑战性的生物系统进行表征成为可能，包括大型DNA结合蛋白和膜蛋白。本方案描述了如何将各种质谱数据（如原生质谱和离子淌度-质谱（IM-MS））与分子动力学模拟相结合，以深入了解解旋酶-核酸酶DNA修复蛋白复合物。由此产生的方法为详细研究配体与参与重要生物过程的其他蛋白质复合物的结合提供了一个框架。
