Alternative fusion protein strategies to express recalcitrant QconCAT proteins for quantitative proteomics of human drug metabolizing enzymes and transporters

Matthew R. Russell, Brahim Achour, Edward A. McKenzie, Ruth Lopez, Matthew D. Harwood, Amin Rostami-Hodjegan, Jill Barber

    Research output: Contribution to journalArticlepeer-review

    Abstract

    QconCAT is a tool for quantitative proteomics, consisting of an artificial protein, expressed from an artificial gene, made up of a concatenated string of proteotypic peptides selected from the proteins under study. Isotopically labeled QconCAT (usually containing 13C6-arginine and 13C6-lysine) provides a standard for each proteotypic peptide included in its sequence. In practice, some QconCAT proteins fail to express at sufficient levels for the purpose of quantitative analysis. Two complementary methods are presented to express recalcitrant QconCAT proteins intended to quantify human hepatic enzymes and transporters. © 2013 American Chemical Society.
    Original languageEnglish
    Pages (from-to)5934-5942
    Number of pages8
    JournalJournal of Proteome Research
    Volume12
    Issue number12
    DOIs
    Publication statusPublished - 6 Dec 2013

    Keywords

    • fusion construct
    • insoluble fusion expression protein
    • MetCAT
    • NUSa-soluble fusion expression protein
    • SDS-PAGE
    • TransCAT
    • triple-quadrupole mass spectrometry

    Research Beacons, Institutes and Platforms

    • Manchester Institute of Biotechnology

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