Quantitative Proteomics Employing Primary Amine Affinity Tags |
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| Authors: | Van M. Hoang Thomas P. Conrads Timothy D. Veenstra Josip Blonder Atsushi Terunuma Jonathan C. Vogel Robert J. Fisher |
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| Affiliation: | aProtein Chemistry Laboratory.;bMass Spectrometry Center.;cBiomedical Proteomics Program, National Cancer Institute at Frederick, SAIC-Frederick, Frederick, MD; ;dDermatology Branch, National Cancer Institute, Bethesda, MD |
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| Abstract: | A proteomics-based method using stable isotope labeling to assess the relative abundance of peptides or proteins is described. Bradykinin and carbonic anhydrase were labeled with sulfosuccinimidyl-2-(biotinamido) ethyl-1,3-dithiopropionate, a membrane impermeant reagent that is reactive with primary amines. Specificity of the label to primary amines was demonstrated using tandem mass spectrometry. Also, relative quantitation was achieved by secondary labeling with natural isotopic abundance and stable isotope-labeled methyl iodide. We believe this to be an effective stable isotope-labeling method for quantitative proteomics. |
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| Keywords: | stable isotope labeling Sulfo-NHS-SS-biotin proteomics |
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