on 06-05-201207:34 AM - edited on 10-15-202111:01 AM by Closed Account
Bomgarden RD, Viner RI, Kuhn K, Pike I, Rogers JC. ASMS 2012 Poster Purpose: To develop an iodoacetyl Tandem Mass Tag (iodoTMT) reagent for irreversible cysteine peptide labeling, enrichment and multiplexed quantitation.
Methods: Reduced sulfhydryls of protein cysteines were labeled with iodoTMTzero and/or iodoTMTsixplex reagents. Labeled peptides were enriched using an immobilized anti-TMT antibody resin before mass spectrometry (MS) analysis.
Results:We developed an iodoTMT reagent set to perform duplex isotopic or sixplex isobaric mass spectrometry (MS) quantitation of cysteine-containing peptides. IodoTMT reagents showed efficient and specific labeling of peptide cysteine residues with reactivity similar to iodoacetamide. Using an anti-TMT antibody, we characterized the immunoaffinity enrichment of peptides labeled with iodoTMT reagents from complex protein cell lysates and for detection of S-nitrosylated cysteines.