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• W2122103660 endingPage "593" @default.
• W2122103660 startingPage "577" @default.
• W2122103660 abstract "An amine specific peptide derivatization strategy involving the use of novel isobaric stable isotope encoded ‘fixed charge’ sulfonium ion reagents, coupled with an analysis strategy employing capillary HPLC, ESI-MS, and automated data dependent ion trap CID-MS/MS, -MS3, and/or ETD-MS/MS, has been developed for the improved quantitative analysis of protein phosphorylation, and for identification and characterization of their site(s) of modification. Derivatization of 50 synthetic phosphopeptides with S,S′-dimethylthiobutanoylhydroxysuccinimide ester iodide (DMBNHS), followed by analysis using capillary HPLC-ESI-MS, yielded an average 2.5-fold increase in ionization efficiencies and a significant increase in the presence and/or abundance of higher charge state precursor ions compared to the non-derivatized phosphopeptides. Notably, 44% of the phosphopeptides (22 of 50) in their underivatized states yielded precursor ions whose maximum charge states corresponded to +2, while only 8% (4 of 50) remained at this maximum charge state following DMBNHS derivatization. Quantitative analysis was achieved by measuring the abundances of the diagnostic product ions corresponding to the neutral losses of ‘light’ (S(CH3)2) and ‘heavy’ (S(CD3)2) dimethylsulfide exclusively formed upon CID-MS/MS of isobaric stable isotope labeled forms of the DMBNHS derivatized phosphopeptides. Under these conditions, the phosphate group stayed intact. Access for a greater number of peptides to provide enhanced phosphopeptide sequence identification and phosphorylation site characterization was achieved via automated data-dependent CID-MS3 or ETD-MS/MS analysis due to the formation of the higher charge state precursor ions. Importantly, improved sequence coverage was observed using ETD-MS/MS following introduction of the sulfonium ion fixed charge, but with no detrimental effects on ETD fragmentation efficiency." @default.
• W2122103660 created "2016-06-24" @default.
• W2122103660 creator A5022203538 @default.
• W2122103660 creator A5024938330 @default.
• W2122103660 creator A5049983845 @default.
• W2122103660 creator A5057698507 @default.
• W2122103660 date "2011-07-06" @default.
• W2122103660 modified "2023-09-27" @default.
• W2122103660 title "Sulfonium Ion Derivatization, Isobaric Stable Isotope Labeling and Data Dependent CID- and ETD-MS/MS for Enhanced Phosphopeptide Quantitation, Identification and Phosphorylation Site Characterization" @default.
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• W2122103660 doi "https://doi.org/10.1007/s13361-011-0190-0" @default.
• W2122103660 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4228788" @default.
• W2122103660 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/21952753" @default.
• W2122103660 hasPublicationYear "2011" @default.
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