Characteristic mass spectral fragments of the organophosphorus agent FP-biotin and FP-biotinylated peptides from trypsin and bovine albumin (Tyr410)

Lawrence M Schopfer, Matthew M. Champion, Nate Tamblyn, Charles M. Thompson, Oksana Lockridge

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

A mass spectrometry-based method was developed for selective detection of FP-biotinylated peptides in complex mixtures. Mixtures of peptides, at the low-picomole level, were analyzed by liquid chromatography and positive ion, nanospray, triple quadrupole, linear ion trap mass spectrometry. Peptides were fragmented by collision-activated dissociation in the mass spectrometer. The free FP-biotin and peptides containing FP-biotinylated serine or FP-biotinylated tyrosine yielded characteristic fragment ions at 227, 312, and 329 m/z. FP-biotinylated serine yielded an additional characteristic fragment ion at 591 m/z. Chromatographic peaks containing FP-biotinylated peptides were indicated by these diagnostic ions. Data illustrating the selectivity of the approach are presented for tryptic digests of FP-biotinylated trypsin and FP-biotinylated serum albumin. A 16-residue peptide from bovine trypsin was biotinylated on the active site serine. A 3-residue peptide from bovine albumin, YTR, was biotinylated on Tyr410. This latter result confirms that the organophosphorus binding site of albumin is a tyrosine. This method can be used to search for new biomarkers of organophosphorus agent exposure.

Original languageEnglish (US)
Pages (from-to)122-132
Number of pages11
JournalAnalytical Biochemistry
Volume345
Issue number1
DOIs
StatePublished - Oct 1 2005

Fingerprint

Trypsin
Albumins
Peptides
Ions
Serine
Mass spectrometry
Tyrosine
Mass Spectrometry
Liquid chromatography
Biomarkers
Mass spectrometers
10-(fluoroethoxyphosphinyl)-N-(biotinamidopentyl)decanamide
Complex Mixtures
Serum Albumin
Liquid Chromatography
Catalytic Domain
Positive ions
Binding Sites

Keywords

  • Albumin
  • Characteristic ions
  • FP-biotin
  • Quadrupole mass spectrometry
  • Trypsin

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Characteristic mass spectral fragments of the organophosphorus agent FP-biotin and FP-biotinylated peptides from trypsin and bovine albumin (Tyr410). / Schopfer, Lawrence M; Champion, Matthew M.; Tamblyn, Nate; Thompson, Charles M.; Lockridge, Oksana.

In: Analytical Biochemistry, Vol. 345, No. 1, 01.10.2005, p. 122-132.

Research output: Contribution to journalArticle

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