title: Application of electrostatic repulsion hydrophilic interaction chromatography to the characterization of proteome, glycoproteome, and phosphoproteome using nano LC-MS/MS.
abstract: In shotgun proteomics, peptide fractionation is essential for in-depth characterization of proteomes and the mapping of protein posttranslational modifications. Recently, a mix-mode chromatography [i.e., electrostatic repulsion hydrophilic interaction chromatography (ERLIC)] has been developed and found to be a versatile method in proteome characterization. Here, we use ERLIC to characterize the glycoproteome and phosphoproteome simultaneously. We also demonstrate that the ERLIC can be an alternative to the commonly used strong cation exchange chromatography for higher recovery of proteins during whole proteome analysis. These protocols can be easily adopted in most proteomics laboratories.
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore, Singapore.
authors.forename: Piliang
authors.initials: P
authors.lastname: Hao
authors.forename: Huoming
authors.initials: H
authors.lastname: Zhang
authors.forename: Siu Kwan
authors.initials: SK
authors.lastname: Sze
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D057927:Hydrophobic and Hydrophilic Interactions; D051379:Mice; D036103:Nanotechnology / Q000379:methods*; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D020543:Proteome / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D051381:Rats; D055672:Static Electricity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D020543:Proteome; D014357:Trypsin