Automating mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan

J Proteomics. 2015 Sep 8;127(Pt A):211-22. doi: 10.1016/j.jprot.2015.05.015. Epub 2015 May 21.

Abstract

Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. Hence, the growing importance of glycoproteomics necessitates in-depth qualitative and quantitative studies of glycans. One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. Recently, we have introduced aminoxyTMT reagents, which enable efficient relative quantitation of carbohydrates, improved glycan ionization efficiency and increased analytical throughput. These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. This analysis typically includes identification/quantitation of glycans using MS/MS data and differential analysis across biological samples. We have developed software modules to streamline such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures. This article is part of a Special Issue entitled: Proteomics in India.

Keywords: AminoxyTMT 6; Glycan quantitation; Reporter ions; SimGlycan; Tandem mass spectrometry.

MeSH terms

  • Animals
  • Cattle
  • Glycomics / methods*
  • Glycoproteins / analysis*
  • Glycosylation
  • Mass Spectrometry / methods*

Substances

  • Glycoproteins