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• W2009522270 abstract "The ubiquitously expressed reduced folate carrier (RFC) is the major transport system for folate cofactors in mammalian cells and tissues. Previous considerations of RFC structure and mechanism were based on the notion that RFC monomers were sufficient to mediate transport of folate and antifolate substrates. The present study examines the possibility that human RFC (hRFC) exists as higher order homo-oligomers. By chemical cross-linking, transiently expressed hRFC in hRFC-null HeLa (R5) cells with the homobifunctional cross-linker 1,3-propanediyl bis-methanethiosulfonate and Western blotting, hRFC species with molecular masses of hRFC homo-oligomers were identified. Hemagglutinin- and Myc epitope-tagged hRFC proteins expressed in R5 cells were co-immunoprecipitated from both membrane particulate and surface-enriched membrane fractions, indicating that oligomeric hRFC is expressed at the cell surface. By co-expression of wild type and inactive mutant S138C hRFCs, combined with surface biotinylation and confocal microscopy, a dominant-negative phenotype was demonstrated involving greatly decreased cell surface expression of both mutant and wild type carrier caused by impaired intracellular trafficking. For another hRFC mutant (R373A), expression of oligomeric wild type-mutant hRFC was accompanied by a significant and disproportionate loss of wild type activity unrelated to the level of surface carrier. Collectively, our results demonstrate the existence of hRFC homo-oligomers. They also establish the likely importance of these higher order hRFC structures to intracellular trafficking and carrier function." @default.
• W2009522270 created "2016-06-24" @default.
• W2009522270 creator A5017584174 @default.
• W2009522270 creator A5066746739 @default.
• W2009522270 date "2009-01-01" @default.
• W2009522270 modified "2023-10-18" @default.
• W2009522270 title "Oligomeric Structure of the Human Reduced Folate Carrier" @default.
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• W2009522270 doi "https://doi.org/10.1074/jbc.m807206200" @default.
• W2009522270 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2631974" @default.
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• W2009522270 hasPublicationYear "2009" @default.
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