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).

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2013–2014

“…The product was fragmented under low‐energy CID conditions to yield a simplified ion series of both the glycan and the peptide that could be used for sequencing. The method was applied to glycopeptides from α 1‐acid glycoprotein and both the glycan and peptide in each glycopeptide were successfully sequenced by MALDI‐MS/MS (Taniguchi et al, ).…”

Relative Quantitation of Glycopeptides Based on Stable Isotope Labeling Using MALDI-TOF MS

“…This work describes how we developed a novel strategy based on stable isotope labeling allowing the highly sensitive relative quantitation of glycopeptides. The labeling of glycopeptides has been applied to cation-charged mass tags [35] bearing a quaternary phosphonium cation or a tertiary, quaternary ammonium cation, such as tris-(2,4,6-trimethoxyphenyl)-phosphonium (TMPP) derivatives, and also to a tandem mass tag (TMT) [36,37]. These approaches are similar to those used in proteomics and glycomics [38].…”

Quantitative Metabolite Profiling of an Amino Group Containing Pharmaceutical in Human Plasma via Precolumn Derivatization and High-Performance Liquid Chromatography-Inductively Coupled Plasma Mass Spectrometry

“…Amino functional groups are found in a large fraction of the currently used drug substances, thus the utilization of the reactivity of this functional group offers an excellent opportunity for the development of a potentially generally applicable derivatization procedure, enabling HPLC-ICPMS-based quantitative metabolite profiling of amino group containing pharmaceutical drugs via the introduction of heteroelements into the molecules of interest. Several reagents, such as dansyl chloride, − o-phthalaldehyde, − and N-succinimidyl analogues, − are widely used for the derivatization of amino groups for different purposes. These reagents react rapidly with amino groups under mild conditions and usually, a high yield of derivatives is obtained.…”