title: Comparison and optimization of strategies for a more profound profiling of the sialylated N-glycoproteomics in human plasma using metal oxide enrichment.
abstract: Glycosylation is an important posttranslational modification of proteins and plays a crucial role in both cellular functions and secretory pathways. Sialic acids (SAs), a family of nine-carbon-containing acidic monosaccharides, often terminate the glycan structures of cell surface molecules and secreted glycoproteins and perform an important role in many biological processes. Hence, a more profound profiling of the sialylated glycoproteomics may improve our knowledge of this modification and its effects on protein functions. Here, we systematically investigated different strategies to enrich the SA proteins in human plasma using a newly developed technology that utilizes titanium dioxide for sialylated N-glycoproteomics profiling by mass spectrometry. Our results showed that using a combination of a filter-aided sample preparation method, TiO2 chromatography, multiple enzyme digestion, and two-dimensional reversed-phase peptide fractionation led to a more profound profiling of the SA proteome. In total, 982 glycosylation sites in 413 proteins were identified, among which 37.8% were newly identified, to establish the largest database of sialic acid containing proteins from human plasma.
authors.affiliation: College of Life Science and Bio-engineering, Beijing University of Technology, Beijing 100022, China.
authors.forename: Xinyuan
authors.initials: X
authors.lastname: Zhao
authors.forename: Cheng
authors.initials: C
authors.lastname: Ma
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Han
authors.forename: Jing
authors.initials: J
authors.lastname: Jiang
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.forename: Jifeng
authors.initials: J
authors.lastname: Wang
authors.forename: Wantao
authors.initials: W
authors.lastname: Ying
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Qian
mesh_terms: D005591:Chemical Fractionation / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D006023:Glycoproteins / Q000032:analysis / Q000097:blood* / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry; D014025:Titanium / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry
chemical_list: D006023:Glycoproteins; C009495:titanium dioxide; D014025:Titanium; D014357:Trypsin; D019158:N-Acetylneuraminic Acid