Element-Coded Affinity Tags for Peptides and Proteins
| Autor: | Todd M. Corneillie, Claude F. Meares, Nathaniel G. Butlin, Paul A. Whetstone |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Affinity label
Biomedical Engineering Pharmaceutical Science Bioengineering Peptide Mass spectrometry Tandem mass spectrometry Chromatography Affinity Gas Chromatography-Mass Spectrometry Mass Spectrometry Heterocyclic Compounds 1-Ring chemistry.chemical_compound Affinity chromatography Organometallic Compounds DOTA Trypsin Yttrium Terbium Chromatography High Pressure Liquid Chelating Agents Pharmacology chemistry.chemical_classification Chromatography biology Chemistry Hydrolysis Organic Chemistry Proteins Affinity Labels Elements Combinatorial chemistry biology.protein Indicators and Reagents Metals Rare Earth Monoisotopic mass Peptides Biotechnology Cysteine |
| Zdroj: | Bioconjugate Chemistry. 15:3-6 |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/bc034150l |
| Popis: | Isotope-coded affinity tags (ICAT) represent an important new tool for the analysis of complex mixtures of proteins in living systems [Aebersold, R., and Mann, M. (2003) Nature, 422, 198-207]. We envisage an alternative protein-labeling technique based on tagging with different element-coded metal chelates, which affords affinity chromatography, quantification, and identification of a tagged peptide from a complex mixture. As proof of concept, a synthetic peptide was modified at a cysteine side chain with either a carboxymethyl group or acetamidobenzyl-1,4,7,10-tetraazacyclododecane-N,N',N' ',N' "-tetraacetic acid (AcBD) chelates of terbium or yttrium. A mixture of the three modified peptides in a mole ratio of 100:1.0:0.83 carboxymethyl:AcBD-Tb:AcBD-Y was trypsinized, purified on a new affinity column that binds rare-earth DOTA chelates, and analyzed by LC-MS/MS. Chelate-tagged tryptic peptides eluted cleanly from the affinity column; the tagged peptides chromatographically coeluted during LC-MS analysis, were present in the expected ratio as indicated by MS ion intensity, and were sequence-identified by tandem mass spectrometry. DOTA-rare earth chelates have exceptional properties for use as affinity tags. They are highly polar and water-soluble. Many of the rare earth elements are naturally monoisotopic, providing a variety of simple choices for preparing mass tags. Further, the rare earths are heavy elements, whose mass defects give the masses of tagged peptides exact values not normally shared by molecules that contain only light elements. |
| Databáze: | OpenAIRE |
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