O'Brien Darragh P, Brier Sébastien, Ladant Daniel, Durand Dominique, Chenal Alexandre, Vachette Patrice
Institut Pasteur, UMR CNRS 3528, Chemistry and Structural Biology Department, Paris, France.
Institut Pasteur, USR CNRS 2000, CITECH, Chemistry and Structural Biology Department, Paris, France.
Biotechnol Appl Biochem. 2018 Jan;65(1):62-68. doi: 10.1002/bab.1577. Epub 2017 Oct 2.
Small-angle X-ray scattering (SAXS) is a relatively simple experimental technique that provides information on the global conformation of macromolecules in solution, be they fully structured, partially, or extensively unfolded. Size exclusion chromatography in line with a SAXS measuring cell considerably improves the monodispersity and ideality of solutions, the two main requirements of a "good" SAXS sample. Hydrogen/deuterium exchange monitored by mass spectrometry (HDX-MS) offers a wealth of information regarding the solvent accessibility at the local (peptide) level. It constitutes a sensitive probe of local flexibility and, more generally, of structural dynamics. The combination of both approaches presented here is very powerful, as illustrated by the case of RD, a calcium-binding protein that is part of a bacterial virulence factor.
小角X射线散射（SAXS）是一种相对简单的实验技术，可提供溶液中大分子整体构象的信息，无论其是完全结构化、部分展开还是广泛展开。与SAXS测量池联用的尺寸排阻色谱法可显著提高溶液的单分散性和理想性，这是“优质”SAXS样品的两个主要要求。通过质谱监测的氢/氘交换（HDX-MS）可提供有关局部（肽）水平溶剂可及性的丰富信息。它是局部柔韧性以及更普遍的结构动力学的灵敏探针。此处介绍的两种方法相结合非常有效，如作为细菌毒力因子一部分的钙结合蛋白RD的例子所示。
