| DOI | Trouver le DOI : https://doi.org/10.1021/ac070091j |
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| Auteur | Rechercher : Liu, X.; Rechercher : Li, X.; Rechercher : Chan, K.; Rechercher : Zou, Wei; Rechercher : Pribil, P.; Rechercher : Sawyer, M.; Rechercher : Li, J. |
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| Format | Texte, Article |
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| Sujet | acid; asparagine; chromatography; digestion; esterification; form; glycoprotein; glycosylation; laser; mass spectrometry; methyl; methylation; mixture; model; N-linked; ovalbumin; polysaccharides; pronase; protein glycosylation; residues; serum; sialic acids; tandem mass spectrometry |
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| Résumé | A 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. |
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| Date de publication | 2007-05-15 |
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| Maison d’édition | American Chemical Society |
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| Dans | |
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| Langue | anglais |
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| Publications évaluées par des pairs | Oui |
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| Numéro du CNRC | LIU2007A |
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| Numéro NPARC | 9368888 |
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| Exporter la notice | Exporter en format RIS |
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| Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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| Identificateur de l’enregistrement | c9138930-c417-40c4-937f-c82cc61f7672 |
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| Enregistrement créé | 2009-07-10 |
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| Enregistrement modifié | 2020-05-10 |
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