Predicted multiple selected reaction monitoring to screen activated drug-mediated modifications on human serum albumin

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11 Citations (Scopus)

Abstract

Metabolic activation of drugs frequently generates electrophilic products that may undergo covalent binding to biological macromolecules, such as proteins and DNA. The resulting covalent adducts are of considerable concern in drug discovery and development. Several strategies for assessing the potential risks of candidate drugs have been reported. Of these, glutathione trapping is the most commonly used method together with mass spectrometry. Furthermore, drug-mediated protein modifications have been studied using serum albumin and CYP enzymes to clarify target amino acids and mechanism-based inhibition, respectively. In this article, we introduce a practical way to screen drug-mediated protein modifications. The method, referred to as "predicted multiple selected reaction monitoring," is based on the selected reaction monitoring (SRM) strategy, but targets all possible chemically modified tryptic peptides. The creation of SRM lists may require patience; however, this strategy could facilitate more sensitive screening compared with the common strategy of data-dependent product ion scanning. Ketoprofen-N-hydroxysuccinimidyl ester (equivalent to glucuronide) and N-acetyl-p-benzoquinone imine (NAPQI) were allowed to react with human serum albumin as a model experiment. Using this strategy, 11 ketoprofen-adduction sites (at Lys 137, 195, 199, 212, 351, 402, 432, 436, 525, 536, and 541) and 1 NAPQI-adduction site (at Cys34) were easily identified.

Original languageEnglish
Pages (from-to)59-67
Number of pages9
JournalAnalytical Biochemistry
Volume449
Issue number1
DOIs
Publication statusPublished - 2014 Mar 15

Keywords

  • Activated drug
  • Chemical modification
  • Human serum albumin
  • Mass spectrometry
  • Predicted multiple selected reaction monitoring
  • Screening

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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