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http://dx.doi.org/10.5478/MSL.2012.3.2.39

An Application of Electrostatic Repulsion Hydrophilic Interaction Chromatography in Phospho- and Glycoproteome Profiling of Epicardial Adipose Tissue in Obesity Mouse  

Tran, Trang Huyen (Lee Gil Ya Cancer and Diabetes Institute, Gachon University)
Hwang, In-Jae (Institute of Molecular Biology and Genetics School of Biological Sciences, Seoul National University)
Park, Jong-Moon (Lee Gil Ya Cancer and Diabetes Institute, Gachon University)
Kim, Jae-Bum (Institute of Molecular Biology and Genetics School of Biological Sciences, Seoul National University)
Lee, Hoo-Keun (Lee Gil Ya Cancer and Diabetes Institute, Gachon University)
Publication Information
Mass Spectrometry Letters / v.3, no.2, 2012 , pp. 39-42 More about this Journal
Abstract
Phosphorylation and glycosylation are two of the most important and widespread post-translational modifications (PTMs) in an organism. Proteomics analysis of the PTMs has been challenged by low stoichiometry of the modified proteins and suppression effects by high abundance proteins, typically no-functional house-keeping proteins. In this study, a novel method was applied for not only isolating PTM peptides from intact peptides but also concurrently characterizing of glyco- and phosphoproteome using electrostatic repulsion hydrophilic interaction chromatography (ERLIC) packed with silica coated by crosslinked polyethyleneimine. For 2 mg tryptic digest of mouse proteome of epicardial adipose tissue with fat diet, 802 N-glycosylated peptides of 316 glycoproteins and 159 phosphorylated peptides of 75 phosphoproteins were identified using HPLC chip/quadrupole time-of-flight (Q-OF) tandem mass spectrometer.
Keywords
ERLIC; Phosphoproteome enrichment; Glycoproteome enrichment; Simultaneous enrichment; PTMs;
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