Element-Coded Affinity Tags for Peptides and Proteins

Paul A. Whetstone, Nathaniel G. Butlin, Todd M. Corneillie, Claude F. Meares

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)3-6
Number of pages4
JournalBioconjugate Chemistry
Volume15
Issue number1
DOIs
StatePublished - Jan 2004

Fingerprint

Chemical elements
Peptides
Proteins
Rare earths
Complex Mixtures
Terbium
Affinity chromatography
Yttrium
Tandem Mass Spectrometry
Rare earth elements
Affinity Chromatography
Isotopes
Labeling
Mass spectrometry
Cysteine
Metals
Ions
Light
Defects
Molecules

ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Whetstone, P. A., Butlin, N. G., Corneillie, T. M., & Meares, C. F. (2004). Element-Coded Affinity Tags for Peptides and Proteins. Bioconjugate Chemistry, 15(1), 3-6. https://doi.org/10.1021/bc034150l

Element-Coded Affinity Tags for Peptides and Proteins. / Whetstone, Paul A.; Butlin, Nathaniel G.; Corneillie, Todd M.; Meares, Claude F.

In: Bioconjugate Chemistry, Vol. 15, No. 1, 01.2004, p. 3-6.

Research output: Contribution to journalArticle

Whetstone, PA, Butlin, NG, Corneillie, TM & Meares, CF 2004, 'Element-Coded Affinity Tags for Peptides and Proteins', Bioconjugate Chemistry, vol. 15, no. 1, pp. 3-6. https://doi.org/10.1021/bc034150l
Whetstone PA, Butlin NG, Corneillie TM, Meares CF. Element-Coded Affinity Tags for Peptides and Proteins. Bioconjugate Chemistry. 2004 Jan;15(1):3-6. https://doi.org/10.1021/bc034150l
Whetstone, Paul A. ; Butlin, Nathaniel G. ; Corneillie, Todd M. ; Meares, Claude F. / Element-Coded Affinity Tags for Peptides and Proteins. In: Bioconjugate Chemistry. 2004 ; Vol. 15, No. 1. pp. 3-6.
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