Improved Mass Spectrometric Characterization of Protein Glycosylation Reveals Unusual Glycosylation of Maize-Derived Bovine Trypsin

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• 作者：Hao Zhang ; Richard Y− ; C. Huang ; Pegah R. Jalili ; Janet W. Irungu ; Gordon R. Nicol ; Kevin B. Ray ; Henry W. Rohrs ; Michael L. Gross
• 刊名：Analytical Chemistry
• 出版年：2010
• 出版时间：December 15, 2010
• 年：2010
• 卷：82
• 期：24
• 页码：10095-10101
• 全文大小：372K
• 年卷期：v.82,no.24(December 15, 2010)
• ISSN：1520-6882

文摘

Although bottom-up proteomics using tryptic digests is widely used to locate post-translational modifications (PTM) in proteins, there are cases where the protein has several potential modification sites within a tryptic fragment and MS² strategies fail to pinpoint the location. We report here a method using two proteolytic enzymes, trypsin and pepsin, in combination followed by tandem mass spectrometric analysis to provide fragments that allow one to locate the modification sites. We used this strategy to find a glycosylation site on bovine trypsin expressed in maize (TrypZean). Several glycans are present, and all are attached to a nonconsensus N-glycosylation site on the protein.