title: Synthetic hydrophobic peptides are substrates for P-glycoprotein and stimulate drug transport.
abstract: P-Glycoprotein functions as an ATP-driven active efflux pump for many natural products and chemotherapeutic drugs. Hydrophobic peptides have been shown to block drug uptake by P-glycoprotein, indicating that they might be transport substrates. The present study examines the interaction of the synthetic peptide series NAc-LnY-amide with the multidrug transporter. Several peptides in this series caused up to 3.5-fold enhancement of colchicine accumulation in membrane vesicles from multidrug resistant (MDR) cells, which suggests the existence of novel interactions between the binding sites for peptides and drug. Peptides did not stimulate vinblastine transport, which was inhibited as expected for competing substrates. These peptides displayed modest stimulatory effects on the ATPase activity of P-glycoprotein. None blocked azidopine photoaffinity labelling, showing that they probably occupy a binding site separate from that for the drug. Studies with 125I-labelled NAc-LLY-amide showed that it was transported by P-glycoprotein in both membrane vesicles and reconstituted proteoliposomes. Uptake of the peptide was rapid, saturable, osmotically sensitive and occurred against a concentration gradient. The enhancing effect of NAc-LLY-amide on colchicine transport was reciprocated, i.e. colchicine greatly increased the transport of labelled peptide by P-glycoprotein. Peptide transport was also modulated, both positively and negatively, by other MDR spectrum drugs. It is concluded that linear hydrophobic peptides are indeed transported by P-glycoprotein, and some have interactions with drug substrates that result in mutual stimulation of transport.
authors.affiliation: Guelph-Waterloo Centre for Graduate Work in Chemistry, Department of Chemistry and Biochemistry, University of Guelph, Ontario, Canada.
authors.forename: F J
authors.initials: FJ
authors.lastname: Sharom
authors.forename: X
authors.initials: X
authors.lastname: Yu
authors.forename: G
authors.initials: G
authors.lastname: DiDiodato
authors.forename: J W
authors.initials: JW
authors.lastname: Chu
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000345:Affinity Labels; D000818:Animals; D001386:Azides; D001692:Biological Transport / Q000187:drug effects; D016466:CHO Cells; D003078:Colchicine / Q000378:metabolism / Q000633:toxicity; D006224:Cricetinae; D004095:Dihydropyridines; D018432:Drug Resistance, Multiple; D007700:Kinetics; D007976:Leupeptins / Q000494:pharmacology; D009842:Oligopeptides / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology*; D013379:Substrate Specificity; D014747:Vinblastine / Q000378:metabolism
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000345:Affinity Labels; D001386:Azides; D004095:Dihydropyridines; D007976:Leupeptins; D009842:Oligopeptides; D014747:Vinblastine; C041130:azidopine; C032854:leupeptin; D003078:Colchicine