Tracking hydrogen/deuterium exchange at glycan sites in glycoproteins by mass spectrometry.

ABSTRACT: Hydrogen/deuterium exchange coupled to mass spectrometry (HDX-MS) has emerged as a technique for studying glycoproteins, which are often refractory to classical methods. Glycan chains are generally assumed to exchange protons very rapidly, making them invisible to this technique. Here, we show that under conditions commonly used for HDX-MS, acetamido groups within glycan chains retain a significant amount of deuterium. Using mono- and polysaccharide standards along with glycopeptides from a panel of glycoproteins, we demonstrate that N-acetyl hexosamines, along with modified Asn side chains, are responsible for this effect. Model compounds for sialic acid also displayed similar exchange kinetics, but terminal sialic acids in the context of an entire glycan chain did not contribute to deute

SUBMITTER: Guttman M 

PROVIDER: S-EPMC3184328 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature