Document Detail


Tracking hydrogen/deuterium exchange at glycan sites in glycoproteins by mass spectrometry.
MedLine Citation:
PMID:  21863800     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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 deuterium retention. Furthermore, the presence of sialic acid appears to enhance the exchange rate of the nearby N-acetyl glucosamines. The ability to detect deuterium exchange at the glycan level opens the possibility of applying HDX-MS to monitor glycan interactions and dynamics.
Authors:
M Guttman; M Scian; K K Lee
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2011-09-06
Journal Detail:
Title:  Analytical chemistry     Volume:  83     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2011 Oct 
Date Detail:
Created Date:  2011-09-30     Completed Date:  2012-01-30     Revised Date:  2012-02-07    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  7492-9     Citation Subset:  IM    
Affiliation:
Department of Medicinal Chemistry, University of Washington, Seattle, Washington 98195, USA. mguttman@uw.edu
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MeSH Terms
Descriptor/Qualifier:
Animals
Carbohydrate Conformation
Cattle
Deuterium Exchange Measurement*
Glycoproteins / chemistry*
Humans
Mass Spectrometry
Polysaccharides / chemistry*
Grant Support
ID/Acronym/Agency:
K99 GM080352-01/GM/NIGMS NIH HHS; K99 GM080352-02/GM/NIGMS NIH HHS; P30AI027767/AI/NIAID NIH HHS; R00 GM080352-03/GM/NIGMS NIH HHS; R00 GM080352-04/GM/NIGMS NIH HHS; R00 GM080352-05/GM/NIGMS NIH HHS; R00GM080352/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Glycoproteins; 0/Polysaccharides

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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