New developments in protein structure–function analysis by MS and use of hydrogen–deuterium exchange microfluidics

• Published in: The FEBS journal Vol. 278; no. 20; pp. 3815 - 3821
• Main Authors: Landreh, Michael, Astorga‐Wells, Juan, Johansson, Jan, Bergman, Tomas, Jörnvall, Hans
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.10.2011
• Subjects: Animals; Biochemistry and Molecular Biology; Biokemi och molekylärbiologi; Biophysics; Crystallography; Deuterium - chemistry; deuterium exchange; Deuterium Exchange Measurement; ESI‐MS; gas‐phase interaction; Humans; Hydrogen - chemistry; ion mobility; Ligands; Mass Spectrometry; Medicin och hälsovetenskap; Microfluidics; Models, Molecular; protein structure; Proteins; Proteins - chemistry; Proteins - metabolism; Structure-Activity Relationship
• ISSN: 1742-464X; 1742-4658; 1742-4658
• DOI: 10.1111/j.1742-4658.2011.08215.x

The study of protein structure and function has evolved to become a leading discipline in the biophysical sciences. Although it is not yet possible to determine 3D protein structures from MS data alone, multiple MS‐based techniques can be combined to obtain structural and functional data that are complementary to classical protein structure information obtained from NMR or X‐ray crystallography. Monitoring gas‐phase interactions of noncovalent complexes yields information on binding constants, complex stability, and the nature of interactions. Ion mobility MS and chemical crosslinking strategies can be applied to probe the architecture of macromolecular assemblies and protein–ligand complexes. MS analysis of hydrogen–deuterium exchange can be used to determine the localization of secondary structure elements, binding sites and conformational dynamics of proteins in solution. This minireview focuses first on new strategies that combine these techniques to gain insights into protein structure and function. Using one such strategy, we then demonstrate how a novel hydrogen–deuterium exchange microfluidics tool can be used online with an ESI mass spectrometer to monitor regional accessibility in a peptide, as exemplified with amyloid‐β peptide 1–40. Mass spectrometry (MS) is a powerful tool in protein structure analysis. In this review, we summarize some of the recent developments and applications in gas phase fragmentation, ion mobility technology, chemical crosslinking and hydrogen/deuterium exchange (HDX) mass spectrometry. We also demonstrate the use of a microfluidic cell for on‐line HDX‐MS to monitor aggregation of the amyloid‐β peptide 1‐40