Protein analysis by hydrogen exchange mass spectrometry.
Mass spectrometry has provided a powerful method for monitoring hydrogen exchange of protein backbone amides with deuterium from solvent. In comparison to popular NMR approaches, mass spectrometry has the advantages of higher sensitivity, wider coverage of sequence, and the ability to analyze larger proteins. Proteolytic fragmentation of proteins following the exchange reaction provides moderate structural resolution, in some cases enabling measurements from single amides. The technique has provided new insight into protein-protein and protein-ligand interfaces, as well as conformational changes during protein folding or denaturation. In addition, recent studies illustrate the utility of hydrogen exchange mass spectrometry toward detecting protein motions relevant to allostery, covalent modifications, and enzyme function.
Hoofnagle
Andrew N
AN
Department of Chemistry and Biochemistry University of Colorado, Boulder, Colorado 80309, USA. Andrew.Hoofnagle@uchsc.edu
Resing
Katheryn A
KA
Ahn
Natalie G
NG
D000577:Amides / Q000737:chemistry*; D003903:Deuterium / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006859:Hydrogen / Q000737:chemistry*; D006860:Hydrogen Bonding; D046911:Macromolecular Substances; D013058:Mass Spectrometry / Q000379:methods*; D009038:Motion; D011485:Protein Binding; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D000577:Amides; D046911:Macromolecular Substances; D011506:Proteins; D006859:Hydrogen; D003903:Deuterium