title: Tandem lectin weak affinity chromatography for glycoprotein enrichment.
abstract: In this chapter we describe the application of lectin weak affinity chromatography (LWAC) for the enrichment of peptides modified by O-linked β-N-acetylglucosamine (O-GlcNAc). O-GlcNAc is a single carbohydrate moiety post-translational modification of intracellular proteins. The stoichiometry of the modification is low and identification of the sites of O-GlcNAc attachment is challenging. To map O-GlcNAc sites we use the approach where a protein sample of interest is digested with trypsin and subjected to LWAC, which employs weak interaction between lectin wheat germ agglutinin and O-GlcNAc. Obtained sample is enriched with O-GlcNAc-modified peptides, which can be identified by means of mass spectrometry.
authors.affiliation: Department of Biochemistry and Molecular Biology, Drexel University, Philadelphia, PA, USA.
authors.forename: Zhi Yuan
authors.initials: ZY
authors.lastname: Ma
authors.forename: Yuliya
authors.initials: Y
authors.lastname: Skorobogatko
authors.forename: Keith
authors.initials: K
authors.lastname: Vosseller
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D017743:Equipment Reuse; D006023:Glycoproteins / Q000302:isolation & purification* / Q000378:metabolism; D013058:Mass Spectrometry; D011138:Polytetrafluoroethylene / Q000737:chemistry; D059748:Proteolysis; D012685:Sepharose / Q000737:chemistry; D014357:Trypsin / Q000378:metabolism; D014909:Wheat Germ Agglutinins / Q000378:metabolism*
chemical_list: D006023:Glycoproteins; D014909:Wheat Germ Agglutinins; D011138:Polytetrafluoroethylene; D012685:Sepharose; D014357:Trypsin; D000117:Acetylglucosamine