An antibiotic linked to peptides and proteins is released by electron capture dissociation fourier transform ion cyclotron resonance mass spectrometry

Clifton K. Fagerquist, Robert R. Hudgins, Mark R. Emmett, Kristina Håkansson, Alan G. Marshall

Research output: Contribution to journalArticle

17 Scopus citations


Desfuroylceftiofur (DFC) is a bioactive β-lactam antibiotic metabolite that has a free thiol group. Previous experiments have shown release of DFC from plasma extracts after addition of a disulfide reducing agent, suggesting that DFC may be bound to plasma and tissue proteins through disulfide bonds. We have reacted DFC with [Arg8]-vasopressin (which has one disulfide bond) and bovine insulin (which has three disulfide bonds) and analyzed the reaction products by use of electron capture dissociation Fourier transform ion cyclotron resonance mass spectrometry (ECD FT-ICR MS), which has previously shown preferential cleavage of disulfide bonds. We observe cleavage of DFC from vasopressin and insulin during ECD, suggesting that DFC is indeed bound to peptides and proteins through disulfide bonds. Specifically, we observed dissociative loss of one, as well as two, DFC species during ECD of [vasopressin + 2(DFC-H) + 2H]2+ from a single electron capture event. Loss of two DFCs could arise from either consecutive or simultaneous loss, but in any case implies a gas phase disulfide exchange step. ECD of [insulin + DFC + 4H]4+ shows preferential dissociative loss of DFC. Combined with HPLC, ECD FT-ICR-MS may be an efficient screening method for detection of drug-biomolecule binding.

Original languageEnglish (US)
Pages (from-to)302-310
Number of pages9
JournalJournal of the American Society for Mass Spectrometry
Issue number4
StatePublished - Apr 2003
Externally publishedYes


ASJC Scopus subject areas

  • Structural Biology
  • Spectroscopy

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