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• W2783192825 endingPage "215" @default.
• W2783192825 startingPage "208" @default.
• W2783192825 abstract "The proton-coupled folate transporter (PCFT) is ubiquitously expressed in solid tumors to which it delivers antifolates, particularly pemetrexed, into cancer cells. Studies of PCFT-mediated transport, to date, have focused exclusively on the influx of folates and antifolates. This article addresses the impact of PCFT on concentrative transport, critical to the formation of the active polyglutamate congeners, and at pH levels relevant to the tumor microenvironment. An HeLa-derived cell line was employed, in which folate-specific transport was mediated exclusively by PCFT. At pH 7.0, there was a substantial chemical gradient for methotrexate that decreased as the extracellular pH was increased. A chemical gradient was still detected at pH 7.4 in the usual HEPES-based transport buffer in contrast to what was observed in a bicarbonate/CO₂-buffered medium. This antifolate gradient correlated with an alkaline intracellular pH in the former (pH 7.85), but not the latter (pH 7.39), buffer and was abolished by the protonophore carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone. The gradient in HEPES buffer at pH 7.4 was the result of the activity of Na⁺/H⁺ exchanger(s); it was eliminated by inhibitors of Na⁺/H⁺ exchanger (s) or Na⁺/K⁺ ATPase. An antifolate chemical gradient was also detected in bicarbonate buffer at pH 6.9 versus 7.4, also suppressed by carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone. When the membrane potential is considered, PCFT generates substantial transmembrane electrochemical-potential gradients at extracellular pH levels relevant to the tumor microenvironment. The augmentation of intracellular pH, when cells are in a HEPES buffer, should be taken into consideration in studies that encompass all proton-coupled transporter families." @default.
• W2783192825 created "2018-01-26" @default.
• W2783192825 creator A5017337262 @default.
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• W2783192825 date "2018-01-11" @default.
• W2783192825 modified "2023-09-27" @default.
• W2783192825 title "Concentrative Transport of Antifolates Mediated by the Proton-Coupled Folate Transporter (SLC46A1); Augmentation by a HEPES Buffer" @default.
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• W2783192825 doi "https://doi.org/10.1124/mol.117.110445" @default.
• W2783192825 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5801552" @default.
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