SEC‐SAXS and HDX‐MS: A powerful combination. The case of the calcium‐binding domain of a bacterial toxin. (2nd October 2017)

• Record Type: Journal Article
• Authors: O'Brien, Darragh P.; Brier, Sébastien; Ladant, Daniel; Durand, Dominique; Chenal, Alexandre; Vachette, Patrice
• Other Names: Dainese Enrico guestEditor.; Gilardi Gianfranco guestEditor.; Maccarrone Mauro guestEditor.
• Is Part Of: Biotechnology and applied biochemistry. Volume 65:Number 1(2018)
• Page Start: 62
• Page End: 68
• Publication Date: 2017-10-02
• Subjects: size‐exclusion chromatography -- small‐angle X‐ray scattering -- hydrogen/deuterium exchange mass spectrometry -- RTX motif -- calcium; Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Genetic Techniques -- Periodicals; Microbiological Techniques -- Periodicals; 660.6
• DOI: 10.1002/bab.1577
• Languages: English
• ISSNs: 0885-4513

Abstract

Abstract: 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.