ArticleEukaryotic cell2007
Characterization of three classes of membrane proteins involved in fungal azole resistance by functional hyperexpression in Saccharomyces cerevisiae.
Article in Eukaryotic cell, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 98 papers.
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98 citing papers in PubMed, 190 citations in OpenAlex.
- ATP-driven dynamics and itraconazole binding of the drug efflux pump Cdr1 from Candida glabrata.The EMBO journal · 2026Article
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- Multidrug efflux pumps and innate azole resistance of Mucor lusitanicus.The Journal of antimicrobial chemotherapy · 2025Article
- Functional analysis of Candida albicans Cdr1 through homologous and heterologous expression studies.FEMS yeast research · 2025Review
- Use of gain-of-function suppressor screening to elucidateFrontiers in microbiology · 2025Article
- Intrinsic resistance to short-tailed azoles in the basal fungusFrontiers in microbiology · 2025Article
- The Molecular Basis of the Intrinsic and Acquired Resistance to Azole Antifungals inJournal of fungi (Basel, Switzerland) · 2024Article
- Batzelladine D, a Marine Natural Product, Reverses the Fluconazole Resistance Phenotype Mediated by Transmembrane Transporters inMarine drugs · 2024Article
- An in vitro set-up to study Pdr5-mediated substrate translocation.Protein science : a publication of the Protein Society · 2024Article
- Effects of efflux pumps on antifungal activity of chitosan againstJournal of oral microbiology · 2024Article
- Synthesis of Altissimacoumarin D and Other Prenylated Coumarins and Their Ability to Reverse the Multidrug Resistance Phenotype inJournal of fungi (Basel, Switzerland) · 2023Article
- Clorgyline Analogs Synergize with Azoles against Drug Efflux inJournal of fungi (Basel, Switzerland) · 2023Article
- Functional Expression of RecombinantJournal of fungi (Basel, Switzerland) · 2023Article
- ExploringJournal of fungi (Basel, Switzerland) · 2022Article
- Genomic landscape of the DHA1 family in Candida auris and mapping substrate repertoire of CauMdr1.Applied microbiology and biotechnology · 2022Article
- Extracts from Argentinian native plants reverse fluconazole resistance in Candida species by inhibiting the efflux transporters Mdr1 and Cdr1.BMC complementary medicine and therapies · 2022Article
- Article
38 more citing papers are in PubMed but not listed here.
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Authors and funding
9 authors at 2 institutions in 2 countries.
Funding
Abstract
The study of eukaryotic membrane proteins has been hampered by a paucity of systems that achieve consistent high-level functional protein expression. We report the use of a modified membrane protein hyperexpression system to characterize three classes of fungal membrane proteins (ABC transporters Pdr5p, CaCdr1p, CaCdr2p, CgCdr1p, CgPdh1p, CkAbc1p, and CneMdr1p, the major facilitator superfamily transporter CaMdr1p, and the cytochrome P450 enzyme CaErg11p) that contribute to the drug resistance phenotypes of five pathogenic fungi and to express human P glycoprotein (HsAbcb1p). The hyperexpression system consists of a set of plasmids that direct the stable integration of a single copy of the expression cassette at the chromosomal PDR5 locus of a modified host Saccharomyces cerevisiae strain, ADDelta. Overexpression of heterologous proteins at levels of up to 29% of plasma membrane protein was achieved. Membrane proteins were expressed with or without green fluorescent protein (GFP), monomeric red fluorescent protein, His, FLAG/His, Cys, or His/Cys tags. Most GFP-tagged proteins tested were correctly trafficked within the cell, and His-tagged proteins could be affinity purified. Kinetic analysis of ABC transporters indicated that the apparent K(m) value and the V(max) value of ATPase activities were not significantly affected by the addition of His tags. The efflux properties of seven fungal drug pumps were characterized by their substrate specificities and their unique patterns of inhibition by eight xenobiotics that chemosensitized S. cerevisiae strains overexpressing ABC drug pumps to fluconazole. The modified hyperexpression system has wide application for the study of eukaryotic membrane proteins and could also be used in the pharmaceutical industry for drug screening.
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