Quantitative proteomics employing primary amine affinity tags.
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.
D000588:Amines / Q000378:metabolism*; D000595:Amino Acid Sequence; D002256:Carbonic Anhydrases / Q000235:genetics; D008239:Lysine / Q000378:metabolism*; D008969:Molecular Sequence Data; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013194:Staining and Labeling / Q000379:methods*
D000588:Amines; D002256:Carbonic Anhydrases; D008239:Lysine