Minimizing back exchange in the hydrogen exchange-mass spectrometry experiment.

ABSTRACT: The addition of mass spectrometry (MS) analysis to the hydrogen exchange (HX) proteolytic fragmentation experiment extends powerful HX methodology to the study of large biologically important proteins. A persistent problem is the degradation of HX information due to back exchange of deuterium label during the fragmentation-separation process needed to prepare samples for MS measurement. This paper reports a systematic analysis of the factors that influence back exchange (solution pH, ionic strength, desolvation temperature, LC column interaction, flow rates, system volume). The many peptides exhibit a range of back exchange due to intrinsic amino acid HX rate differences. Accordingly, large back exchange leads to large variability in D-recovery from one residue to another as well as one pe

SUBMITTER: Walters BT 

PROVIDER: S-EPMC3515739 | biostudies-literature | 2012 Dec

REPOSITORIES: biostudies-literature