Use of a yeast-based membrane protein expression technology to overexpress drug resistance efflux pumps.
Azole antifungal drugs are used widely to treat people with oral fungal infections. Unfortunately, fungi can develop resistance to these drugs. This resistance can be due to the overexpression or mutation of cytochrome P450 14alpha-lanosterol demethylase, also known as ERG11 or CYP51, and/or the overexpression of membrane-located multidrug efflux pumps. We have developed a heterologous membrane protein expression system that can be used to study the structure and function of these proteins in the non-pathogenic, genetically stable, and versatile eukaryotic model organism, Saccharomyces cerevisiae. In this chapter we describe the techniques used to express the Candida albicans efflux pump Cdr1p in S. cerevisiae.
Lamping
Erwin
E
Department of Oral Sciences, School of Dentistry, University of Otago, Dunedin, New Zealand.
Cannon
Richard D
RD
D018528:ATP-Binding Cassette Transporters; D000935:Antifungal Agents / Q000378:metabolism / Q000494:pharmacology; D001393:Azoles / Q000378:metabolism / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics / Q000502:physiology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008957:Models, Genetic; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
D018528:ATP-Binding Cassette Transporters; D000935:Antifungal Agents; D001393:Azoles; D005656:Fungal Proteins; D008565:Membrane Proteins; D029701:Saccharomyces cerevisiae Proteins; D003577:Cytochrome P-450 Enzyme System; C526348:Erg11 protein, S cerevisiae