title: Strategy for glycoproteomics: identification of glyco-alteration using multiple glycan profiling tools.
abstract: Glycan alterations of proteins, a common feature of cancer cells, are associated with carcinogenesis, invasion and metastasis. Glycomics, the study of glycans and glycan-binding proteins in various biological systems, is an emerging field in the postgenome and postproteomics era. However, systematic and robust strategies for glycomics are still not fully established because the structural analysis of glycans, which comprise different patterns of branching, various possible linkage positions as well as monomer anomericity, is technically difficult. Here, we introduce a new strategy for glyco-alteration analysis of glycoproteins by using multiple glycan profiling tools. To understand glycan alterations of proteins by correlating the glycosyltransferase expression profile with the actual glycan structure, we systematically used three glycan profiling tools: (1) multiplex quantitative PCR (qPCR) array format for profiling the expression pattern of glycogenes, (2) lectin microarray as a multiplex glycan-lectin interaction analysis system for profiling either a pool of cell glycoproteins or a target glycoprotein, and (3) tandem mass spectrometry for identifying the glycan structure connected to a target glycoprotein. Using our system, we successfully identified glycan alterations on alpha-fetoprotein (AFP), including a novel LacdiNAc structure in addition to previously reported alterations such as alpha1,6 fucosylation.
authors.affiliation: Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Umezono, Tsukuba, Ibaraki 305-8568, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Ito
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kuno
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Sawaki
authors.forename: Maki
authors.initials: M
authors.lastname: Sogabe
authors.forename: Hidenori
authors.initials: H
authors.lastname: Ozaki
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Masashi
authors.initials: M
authors.lastname: Mizokami
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Shoda
authors.forename: Takashi
authors.initials: T
authors.lastname: Angata
authors.forename: Takashi
authors.initials: T
authors.lastname: Sato
authors.forename: Jun
authors.initials: J
authors.lastname: Hirabayashi
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Ikehara
authors.forename: Hisashi
authors.initials: H
authors.lastname: Narimatsu
mesh_terms: D002460:Cell Line; D054794:Glycomics / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D037102:Lectins / Q000032:analysis*; D011134:Polysaccharides / Q000032:analysis*; D053719:Tandem Mass Spectrometry; D000509:alpha-Fetoproteins / Q000378:metabolism*
chemical_list: D006023:Glycoproteins; D037102:Lectins; D011134:Polysaccharides; D000509:alpha-Fetoproteins