Britt Hannah M, Ben-Younis Aisha, Page Nathanael, Thalassinos Konstantinos
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom.
LGC Group, Teddington TW11 0LY, United Kingdom.
J Am Soc Mass Spectrom. 2024 Dec 4;35(12):3203-3213. doi: 10.1021/jasms.4c00361. Epub 2024 Oct 25.
Native top-down mass spectrometry is a powerful approach for characterizing proteoforms and has recently been applied to provide similarly powerful insights into protein conformation. Current approaches, however, are limited such that structural insights can only be obtained for the entire conformational landscape in bulk or without any direct conformational measurement. We report a new ion-mobility-enabled method for performing native top-down MS in a conformation-specific manner. Our approach identified conformation-linked differences in backbone dissociation for the model protein calmodulin, which simultaneously informs upon proteoform variations and provides structural insights. We also illustrate that our method can be applied to protein-ligand complexes, either to identify components or to probe ligand-induced structural changes.
原生自上而下质谱分析法是一种用于表征蛋白质异构体的强大方法，最近已被应用于对蛋白质构象提供同样强大的见解。然而，目前的方法存在局限性，即只能在整体上获得整个构象格局的结构见解，或者无法进行任何直接的构象测量。我们报告了一种新的基于离子淌度的方法，用于以构象特异性方式进行原生自上而下质谱分析。我们的方法识别出了模型蛋白钙调蛋白在主链解离方面与构象相关的差异，这同时揭示了蛋白质异构体的变化并提供了结构见解。我们还表明，我们的方法可应用于蛋白质 - 配体复合物，既可以识别其组成成分，也可以探测配体诱导的结构变化。
