title: Alternative fusion protein strategies to express recalcitrant QconCAT proteins for quantitative proteomics of human drug metabolizing enzymes and transporters.
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 (13)C6-arginine and (13)C6-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.
authors.affiliation: Manchester Pharmacy School, University of Manchester , Stopford Building, Oxford Road, Manchester M13 9PT, United Kingdom.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Russell
authors.forename: Brahim
authors.initials: B
authors.lastname: Achour
authors.forename: Edward A
authors.initials: EA
authors.lastname: Mckenzie
authors.forename: Ruth
authors.initials: R
authors.lastname: Lopez
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Harwood
authors.forename: Amin
authors.initials: A
authors.lastname: Rostami-Hodjegan
authors.forename: Jill
authors.initials: J
authors.lastname: Barber
mesh_terms: D018938:Caco-2 Cells; D002247:Carbon Isotopes; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D005813:Genes, Synthetic; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D008658:Inactivation, Metabolic / Q000235:genetics*; D008099:Liver / Q000737:chemistry* / Q000201:enzymology; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D011107:Polylysine / Q000378:metabolism; D021381:Protein Transport; D040901:Proteomics; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D002247:Carbon Isotopes; D018832:Molecular Chaperones; D010455:Peptides; D011993:Recombinant Fusion Proteins; D011107:Polylysine; C015462:polyarginine