"One-pot" methylation in glycomics application: esterification of sialic acids and permanent charge construction: Anal.Chem.

Download
  1. Get@NRC: "One-pot" methylation in glycomics application: esterification of sialic acids and permanent charge construction: Anal.Chem. (Opens in a new window)
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titleAnal.Chem.
Volume79
Issue10
Pages38943900; # of pages: 7
SubjectACID; Acids; analysis; Asparagine; Canada; chemistry; Chromatography; Digestion; Esterification; Form; glycoprotein; Glycoproteins; Glycosylation; laser; mass spectrometry; MASS-SPECTROMETRY; method; Methods; METHYL; Methylation; Mixture; MODEL; N-linked; Ovalbumin; Polysaccharides; Pronase; protein; PROTEIN GLYCOSYLATION; RESIDUES; Serum; SIALIC; Sialic Acids; SIALIC-ACID; Tandem Mass Spectrometry; technique
AbstractA simple and rapid "one-pot" methylation method to esterify sialic acids and construct a permanent charge was developed for N-linked glycan analysis, which combined complete nonspecific proteolytic digestion and methylation. A mixture of Asn-glycans prepared from Pronase E digestion of the glycoprotein was passed through a cation-exchange column to convert carboxylic acids to the Na+ form before being methylated with methyl iodide. Derivatives could be easily purified with a hydrophilic affinity chromatography cartridge. Mass spectrometry analysis was performed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) and MALDI-TOF/TOF. The mass spectrometric data indicated that carboxylic acids were methylated in addition to the formation of a quaternary ammonium in the amino group of asparagine residues. Three model glycoproteins, including ribonuclease B, ovalbumin, and transferrin, were employed to demonstrate the merits of this technique. Results showed that the stabilization of sialic acid was achieved in addition to the formation of a permanent charge. Compared to the analysis of underivatized N-glycans, detection sensitivity improved approximately 10-fold. The new technique was further evaluated with glycan profiling of serum transferrin and proved to be a sensitive method for the characterizing protein glycosylation
Publication date
LanguageEnglish
AffiliationNRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedNo
NRC numberLIU2007A
NPARC number9368888
Export citationExport as RIS
Report a correctionReport a correction
Record identifierc9138930-c417-40c4-937f-c82cc61f7672
Record created2009-07-10
Record modified2016-05-09
Bookmark and share
  • Share this page with Facebook (Opens in a new window)
  • Share this page with Twitter (Opens in a new window)
  • Share this page with Google+ (Opens in a new window)
  • Share this page with Delicious (Opens in a new window)