Determining the structure and mechanism of the human multidrug resistance P-glycoprotein using cysteine-scanning mutagenesis and thiol-modification techniques.
The multidrug resistance P-glycoprotein is an ATP-dependent drug pump that extrudes a broad range of hydrophobic compounds out of cells. Its physiological role is likely to protect us from exogenous and endogenous toxins. The protein is important because it contributes to the phenomenon of multidrug resistance during AIDS and cancer chemotherapy. We have used cysteine-scanning mutagenesis and thiol-modification techniques to map the topology of the protein, show that both nucleotide-binding domains are essential for activity, examine packing of the transmembrane segments, map the drug-binding site, and show that there is cross-talk between the ATP-binding sites and the transmembrane segments.
Loo
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Department of Medicine and Biochemistry, University of Toronto, Rm. 7342, Medical Sciences Building, 1 King's College Circle, Toronto, Ont., Canada.
Clarke
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D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000737:chemistry / Q000378:metabolism*; D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D018432:Drug Resistance, Multiple; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D017510:Protein Folding
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018528:ATP-Binding Cassette Transporters; D002352:Carrier Proteins; D004220:Disulfides; D047908:Intracellular Signaling Peptides and Proteins; C089765:NUBP1 protein, human; D003545:Cysteine