title: MALDI mass spectrometry-based sequence analysis of arginine-containing glycopeptides: improved fragmentation of glycan and peptide chains by modifying arginine residue.
abstract: This paper describes an improved method for the sequence analysis of Arg-containing glycopeptide by MALDI mass spectrometry (MS). The method uses amino group derivatization (4-aza-6-(2,6-dimethyl-1-piperidinyl)-5-oxohexanoic acid N-succinimidyl ester) and removal (carboxypeptidase B) or modification (peptidylarginine deiminase 4) of the arginine residue of the peptide. The derivatization attaches a basic tertiary amine moiety onto the peptides, and the enzymatic treatment removes or modifies the arginine residue. Fragmentation of the resulting glycopeptide under low-energy collision-induced dissociation yielded a simplified ion series of both the glycan and the peptide that can facilitate their sequencing. The feasibility of the method was studied using α1 -acid glycoprotein-derived N-linked glycopeptides, and glycan and peptide in each glycopeptide were successfully sequenced by MALDI tandem MS (MS/MS).
authors.affiliation: Koichi Tanaka Laboratory of Advanced Science and Technology (KTLAST), Shimadzu Corporation, 1, Nishinokyo-Kuwabaracho Nakagyo-ku, Kyoto, 604-8511, Japan.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Taniguchi
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kuyama
authors.forename: Shigeki
authors.initials: S
authors.lastname: Kajihara
authors.forename: Koichi
authors.initials: K
authors.lastname: Tanaka
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000032:analysis* / Q000737:chemistry; D002956:Citrulline / Q000737:chemistry; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D008956:Models, Chemical; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry; D020539:Sequence Analysis, Protein / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D006020:Glycopeptides; D011134:Polysaccharides; D002956:Citrulline; D001120:Arginine
keywords: MALDI-QIT MS; arginine removal; citrullination; glycopeptide