ArticleBiochimica et biophysica acta2008
N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT).
Article in Biochimica et biophysica acta, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 60 papers.
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Who cites it
60 citing papers in PubMed, 86 citations in OpenAlex.
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- Novel localization of folate transport systems in the murine central nervous system.Fluids and barriers of the CNS · 2022Article
- Hyperosmolality in CHO cell culture: effects on the proteome.Applied microbiology and biotechnology · 2022Article
- The evolving biology of the proton-coupled folate transporter: New insights into regulation, structure, and mechanism.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2022Review
- The Concept of Folic Acid in Health and Disease.Molecules (Basel, Switzerland) · 2021Review
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- Impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter.PloS one · 2021Article
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- Cellular Pharmacodynamics of a Novel Pyrrolo[3,2-Molecular pharmacology · 2020Article
- LRRK2 Kinase Inhibition Rescues Deficits in Lysosome Function Due to HeterozygousFrontiers in neuroscience · 2020Article
- Expression of a recombinant bacterial L-asparaginase in human cells.BMC research notes · 2019Article
- Identification of the amino acid residue responsible for the myricetin sensitivity of human proton-coupled folate transporter.Scientific reports · 2019Article
- Direct, gabapentin-insensitive interaction of a soluble form of the calcium channel subunit αScientific reports · 2019Article
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- The promise and challenges of exploiting the proton-coupled folate transporter for selective therapeutic targeting of cancer.Cancer chemotherapy and pharmacology · 2018Review
- Regulation of Reduced Folate Carrier (RFC) by Vitamin D Receptor at the Blood-Brain Barrier.Molecular pharmaceutics · 2017Article
- Evaluating Commercially Available Antibodies for Rat α7 Nicotinic Acetylcholine Receptors.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2017Article
- Structure/functional aspects of the human riboflavin transporter-3 (American journal of physiology. Cell physiology · 2017Article
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4 authors at 1 institution in 1 country.
Funding
Abstract
The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. On Western blot, HsPCFT migrates as a broad band (~55 kDa), higher than predicted (~50 kDa) in cell lines. Western blot analysis required that membrane preparations not be incubated in the loading buffer above 50 degrees C to avoid aggregation of the protein. Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. Single mutants retained full transport activity; the double mutant retained a majority of activity. Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains.
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