Specificity of drug transport mediated by CaMDR1: a major facilitator of Candida albicans.
CaMDR1 encodes a major facilitator superfamily (MFS) protein in Candida albicans whose expression has been linked to azole resistance and which is frequently encountered in this human pathogenic yeast. In this report we have overexpressed CaMdr1p in Sf9 insect cells and demonstrated for the first time that it can mediate methotrexate (MTX) and fluconazole (FLC) transport. MTX appeared to be a better substrate for CaMdr1p among these two tested drugs. Due to severe toxicity of these drugs to insect cells, further characterization of CaMdr1p as a drug transporter could not be done with this system. Therefore, as an alternative, CaMdr1p and Cdr1p, which is an ABC protein (ATP binding cassette) also involved in azole resistance in C. albicans, were independently expressed in a common hypersensitive host JG436 of Saccharomyces cerevisiae. This allowed a better comparison between the functionality of the two export pumps. We observed that while both FLC and MTX are effluxed by CaMdr1p, MTX appeared to be a poor substrate for Cdr1p. JG436 cells expressing Cdr1p thus conferred resistance to other antifungal drugs but remained hypersensitive to MTX. Since MTX is preferentially transported by CaMdr1p, it can be used for studying the function of this MFS protein.
Kohli
A
A
Membrane Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi 110 067, India.
Gupta
V
V
Krishnamurthy
S
S
Hasnain
S E
SE
Prasad
R
R
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000378:metabolism*; D000935:Antifungal Agents / Q000378:metabolism / Q000494:pharmacology; D000964:Antimetabolites, Antineoplastic / Q000378:metabolism / Q000494:pharmacology; D001665:Binding Sites; D001692:Biological Transport; D002176:Candida albicans / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D026141:Drug Resistance, Multiple, Fungal / Q000502:physiology*; D015725:Fluconazole / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D008727:Methotrexate / Q000378:metabolism / Q000494:pharmacology; D014170:Transformation, Genetic
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000935:Antifungal Agents; D000964:Antimetabolites, Antineoplastic; D015725:Fluconazole; D008727:Methotrexate