期刊
PROTEOMICS
卷 11, 期 21, 页码 4229-4242出版社
WILEY-BLACKWELL
DOI: 10.1002/pmic.201000677
关键词
1-Phenyl-3-methyl-5-pyrazolone; ESI-MS; Glycoproteomics; Nonreductive release; O-Glycan
资金
- National Natural Science Foundation of China [30870548]
- Program for New Century Excellent Talents in Universities [NCET-08-0893]
- Hi-Tech Research and Development Program of China [2006AA02Z146]
A novel one-pot procedure for the nonreductive release of O-linked glycans from glycoproteins and the simultaneous derivatization of released glycans with 1-phenyl-3-methyl-5-pyrazolone (PMP) is described. Unlike the traditional reductive beta-elimination, which produces alditols, this new method employs PMP/ammonia aqueous solution as the reaction medium. The O-glycans are released from glycoproteins and derivatized with PMP nonreductively, specifically, and quantitatively. Samples can be easily purified from ammonia, excess PMP, and peptide residues by evaporation, chloroform extraction, and solid-phase extraction (SPE) column fractionation for HPLC, CE, or MS analysis. The procedure has been elaborated with two purified glycoproteins, porcine stomach mucin and bovine fetuin, and successfully applied to O-glycan profiling of a challenging biological specimen, healthy human plasma. This new procedure has shown methodological significance in O-glycan analysis.
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