Crossing the phase boundary to study protein dynamics and function: combination of amide hydrogen exchange in solution and ion fragmentation in the gas phase.
Protein dynamics are the key to understanding their behavior. The static protein structure alone in most cases is insufficient to describe the vast array of complex functions they perform in vivo. Until recently there were relatively few techniques available to investigate the dynamic nature of these proteins. Mass spectrometry has recently emerged as a powerful biophysical method, capable of providing both structural and dynamic information. By utilizing the labile nature of amide hydrogens as a marker of the backbone dynamics in solution, combined with gas-phase dissociation techniques, we now have a high-resolution tool to locate these exchanging hydrogens within the sequence of the protein and to probe the functional importance of its structural elements. In this paper we describe several applications of these methodologies to illustrate the importance of dynamics to the biological functions of proteins.
Kaltashov
Igor A
IA
Department of Chemistry, University of Massachusetts, Amherst, Massachusetts 01003, USA. kaltashov@chem.umass.edu
Eyles
Stephen J
SJ
D000577:Amides / Q000378:metabolism; D000818:Animals; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D005544:Forecasting; D005740:Gases; D006801:Humans; D006859:Hydrogen / Q000378:metabolism; D007477:Ions; D013058:Mass Spectrometry / Q000379:methods; D010446:Peptide Fragments / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D012996:Solutions; D013329:Structure-Activity Relationship
D000577:Amides; D005740:Gases; D007477:Ions; D010446:Peptide Fragments; D011506:Proteins; D012996:Solutions; D006859:Hydrogen