title: Glycopeptide Site Heterogeneity and Structural Diversity Determined by Combined Lectin Affinity Chromatography/IMS/CID/MS Techniques.
abstract: Glycopeptides from a tryptic digest of chicken ovomucoid were enriched using a simplified lectin affinity chromatography (LAC) platform, and characterized by high-resolution mass spectrometry (MS) as well as ion mobility spectrometry (IMS)-MS. The LAC platform effectively enriched the glycoproteome, from which a total of 117 glycopeptides containing 27 glycan forms were identified for this protein. IMS-MS analysis revealed a high degree of glycopeptide site heterogeneity. Comparison of the IMS distributions of the glycopeptides from different charge states reveals that higher charge states allow more structures to be resolved. Presumably the repulsive interactions between charged sites lead to more open configurations, which are more readily separated compared with the more compact, lower charge state forms of the same groups of species. Combining IMS with collision induced dissociation (CID) made it possible to determine the presence of isomeric glycans and to reconstruct their IMS profiles. This study illustrates a workflow involving hybrid techniques for determining glycopeptide site heterogeneity and evaluating structural diversity of glycans and glycopeptides.
authors.affiliation: Department of Chemistry, Indiana University, 800 Kirkwood Ave., Bloomington, IN, 47405, USA.
authors.forename: Feifei
authors.initials: F
authors.lastname: Zhu
authors.forename: Jonathan C
authors.initials: JC
authors.lastname: Trinidad
authors.forename: David E
authors.initials: DE
authors.lastname: Clemmer
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D006020:Glycopeptides / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D007536:Isomerism; D037102:Lectins / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D006020:Glycopeptides; D037102:Lectins
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