Functional purification of the monocarboxylate transporter of the yeast Candida utilis.
Plasma membranes of the yeast, Candida utilis, were solubilized with octyl-beta-D-glucopyranoside and a fraction enriched in the lactate carrier was obtained with DEAE-Sepharose anion-exchange chromatography, after elution with 0.4 M NaCl. The uptake of lactic acid into proteoliposomes, containing the purified protein fraction and cytochrome c oxidase, was dependent on a proton-motive force and the transport specificity was consistent with the one of C. utilis intact cells. Overall, we have obtained a plasma membrane fraction enriched in the lactate carrier of C. utilis in which the transport properties were preserved. Given the similarities between the lactate transport of C. utilis and the one of mammalian cells, this purified system could be further explored to screen for specific lactate inhibitors, with potential therapeutic applications.
Baltazar
Fátima
F
Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal. fbaltazar@ecsaude.uminho.pt
Cássio
Fernanda
F
Leão
Cecília
C
D001671:Biochemistry / Q000379:methods*; D001692:Biological Transport; D001709:Biotechnology / Q000379:methods*; D002175:Candida / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D004305:Dose-Response Relationship, Drug; D003576:Electron Transport Complex IV / Q000302:isolation & purification; D004983:Ethanolamines / Q000737:chemistry; D007773:Lactates / Q000737:chemistry; D019344:Lactic Acid / Q000737:chemistry; D027501:Monocarboxylic Acid Transporters / Q000302:isolation & purification*; D011522:Protons; D012685:Sepharose / Q000737:chemistry
D004983:Ethanolamines; D007773:Lactates; D027501:Monocarboxylic Acid Transporters; D011522:Protons; D019344:Lactic Acid; D012685:Sepharose; D003576:Electron Transport Complex IV; C007369:2-diethylaminoethanol