Spectroscopic and biophysical approaches for studying the structure and function of the P-glycoprotein multidrug transporter.
Multidrug resistance is a serious obstacle to the successful chemotherapeutic treatment of many human cancers. A major cause of multidrug resistance is the overexpression of a 170-kDa plasma membrane protein, known as P-glycoprotein, which appears to function as an ATP-driven efflux pump with a very broad specificity for hydrophobic drugs, peptides, and natural products. P-Glycoprotein is a member of the ABC superfamily and is proposed to consist of two homologous halves, each comprising six membrane-spanning segments and a cytosolic nucleotide binding domain. In recent years, P-glycoprotein has been purified and functionally reconstituted into lipid bilayers, where it retains both ATPase and drug transport activity. The availability of purified active protein has led to substantial advances in our understanding of the molecular structure and mechanism of action of this unique transporter. This review will focus on the recent application of fluorescence spectroscopy, infra-red spectroscopy, circular dichroism spectroscopy, electron microscopy, and other biophysical techniques to the study of P-glycoprotein structure and function.
Sharom
F J
FJ
Guelph-Waterloo Centre for Graduate Work in Chemistry and Biochemistry, Department of Chemistry and Biochemistry, University of Guelph, ON, Canada. sharom@chembio.uoguelph.ca
Liu
R
R
Romsicki
Y
Y
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000737:chemistry* / Q000302:isolation & purification; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000737:chemistry*; D000251:Adenosine Triphosphatases / Q000378:metabolism; D001703:Biophysics / Q000379:methods*; D002152:Calorimetry, Differential Scanning / Q000379:methods; D002942:Circular Dichroism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007700:Kinetics; D008854:Microscopy, Electron / Q000379:methods; D008954:Models, Biological; D005810:Multigene Family; D013050:Spectrometry, Fluorescence / Q000379:methods; D013055:Spectrophotometry, Infrared / Q000379:methods; D013057:Spectrum Analysis / Q000379:methods*
D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000251:Adenosine Triphosphatases