Journal Article

N-Glycoproteome of E14.Tg2a Mouse Embryonic Stem Cells

Tian, Qiang, Hood, Leroy, Ma, Li, Lorang, Cynthia, Lee, Denis, Alexander, Vinita, Hohmann, Laura J., Sun, Bingyun, Yan, Xiaowei, Chandrasena, Lalangi

2013

Overview

E14.Tg2a mouse embryonic stem (mES) cells are a widely used host in gene trap and gene targeting techniques. Molecular characterization of host cells will provide background information for a better understanding of functions of the knockout genes. Using a highly selective glycopeptide-capture approach but ordinary liquid chromatography coupled mass spectrometry (LC-MS), we characterized the N-glycoproteins of E14.Tg2a cells and analyzed the close relationship between the obtained N-glycoproteome and cell-surface proteomes. Our results provide a global view of cell surface protein molecular properties, in which receptors seem to be much more diverse but lower in abundance than transporters on average. In addition, our results provide a systematic view of the E14.Tg2a N-glycosylation, from which we discovered some striking patterns, including an evolutionarily preserved and maybe functionally selected complementarity between N-glycosylation and the transmembrane structure in protein sequences. We also observed an environmentally influenced N-glycosylation pattern among glycoenzymes and extracellular matrix proteins. We hope that the acquired information enhances our molecular understanding of mES E14.Tg2a as well as the biological roles played by N-glycosylation in cell biology in general.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Analysis / Animals / Biology / Cell Membrane - metabolism / Cell surface / Cells, Cultured / Chemistry / Chromatography / Chromatography, Liquid / Complementarity / Embryo cells / Embryonic stem cells / Embryonic Stem Cells - cytology / Embryonic Stem Cells - metabolism / Endoplasmic reticulum / Extracellular matrix / Gene targeting / Genetic aspects / Genetic engineering / Glycopeptides - analysis / Glycopeptides - metabolism / Glycoproteins / Glycoproteins - analysis / Glycoproteins - metabolism / Glycosylation / Liquid chromatography / Mass spectrometry / Mass spectroscopy / Membrane Proteins - metabolism / Metabolism / Mice / Protein structure / Proteins / Proteome - analysis / Proteomics / Receptor mechanisms / Receptors / Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Stem cell transplantation / Stem cells