{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T14:49:31Z","timestamp":1763563771293},"reference-count":37,"publisher":"American Society of Hematology","issue":"24","content-domain":{"domain":["ashpublications.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2010,12,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>The proton-coupled folate transporter (PCFT; SLC46A1) mediates folate transport into enterocytes in the proximal small intestine; pcft loss-of-function mutations are the basis for hereditary folate malabsorption. The current study explored the roles of Asp residues in PCFT function. A novel, homozygous, loss-of-function mutation, D156Y, was identified in a child of Pakistani origin with hereditary folate malabsorption. Of the 6 other conserved Asp residues, only one, D109, is shown to be required for function. D156Y, along with a variety of other substitutions at this site (Trp, Phe, Val, Asn, or Lys), lacked function due to instability of the PCFT protein. Substantial function was preserved with Glu, Gly, and, to a lesser extent, with Ser, Thr, and Ala substitutions. This correlated with PCFT bio-tinylated at the cell surface. In contrast, all D109 mutants, including D109E, lacked function irrespective of pH (4.5, 5.5, and 7.4) or substrate concentration (0.5-100\u03bcM), despite surface expression comparable to wild-type PCFT. Hence, D156 plays a critical role in PCFT protein stability, and D109, located in the first intracellular loop between the second and third transmembrane domains, is absolutely required for PCFT function.<\/jats:p>","DOI":"10.1182\/blood-2010-06-291237","type":"journal-article","created":{"date-parts":[[2010,8,31]],"date-time":"2010-08-31T01:58:57Z","timestamp":1283219937000},"page":"5162-5169","update-policy":"http:\/\/dx.doi.org\/10.1182\/blood.2019cm0000","source":"Crossref","is-referenced-by-count":51,"title":["Functional roles of aspartate residues of the proton-coupled folate transporter (PCFT-SLC46A1); a D156Y mutation causing hereditary folate malabsorption"],"prefix":"10.1182","volume":"116","author":[{"given":"Daniel Sanghoon","family":"Shin","sequence":"first","affiliation":[{"name":"Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY;"},{"name":"Department of Medicine, Albert Einstein College of Medicine, Bronx, NY;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sang Hee","family":"Min","sequence":"additional","affiliation":[{"name":"Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY;"},{"name":"Department of Medicine, Albert Einstein College of Medicine, Bronx, NY;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura","family":"Russell","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Human Genetics, Montreal Children's Hospital, McGill University Health Center, Montreal, QC;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rongbao","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY;"},{"name":"Department of Medicine, Albert Einstein College of Medicine, Bronx, NY;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andras","family":"Fiser","sequence":"additional","affiliation":[{"name":"Department of Systems and Computational Biology, Albert Einstein College of Medicine, Bronx, NY; and"},{"name":"Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"I. David","family":"Goldman","sequence":"additional","affiliation":[{"name":"Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY;"},{"name":"Department of Medicine, Albert Einstein College of Medicine, Bronx, NY;"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"234","reference":[{"key":"2019111723170437400_B1","doi-asserted-by":"crossref","first-page":"e4","DOI":"10.1017\/S1462399409000969","article-title":"Membrane transporters and folate homeostasis: intestinal absorption and transport into systemic com-partments and tissues.","volume":"11","author":"Zhao","year":"2009","journal-title":"Expert Rev Mol Med"},{"key":"2019111723170437400_B2","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/S0083-6729(08)00405-6","article-title":"Structure and function of the reduced folate carrier: a paradigm of a major facilitator superfamily mammalian nutrient transporter.","volume":"79C","author":"Matherly","year":"2008","journal-title":"Vitam Horm"},{"issue":"1","key":"2019111723170437400_B3","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1007\/s10555-007-9048-0","article-title":"The folate receptor: what does it promise in tissue-targeted therapeutics?","volume":"26","author":"Salazar","year":"2007","journal-title":"Cancer Metast Rev"},{"issue":"8","key":"2019111723170437400_B4","doi-asserted-by":"crossref","first-page":"1085","DOI":"10.1016\/j.addr.2004.01.002","article-title":"A review of folate receptor alpha cycling and 5-methyltetrahydrofolate accumulation with an emphasis on cell models in vitro.","volume":"56","author":"Kamen","year":"2004","journal-title":"Adv Drug Deliv Rev"},{"issue":"5","key":"2019111723170437400_B5","doi-asserted-by":"crossref","first-page":"917","DOI":"10.1016\/j.cell.2006.09.041","article-title":"Identification of an intestinal folate transporter and the molecular basis for hereditary folate malabsorption.","volume":"127","author":"Qiu","year":"2006","journal-title":"Cell"},{"issue":"4","key":"2019111723170437400_B6","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1182\/blood-2007-02-077099","article-title":"The spectrum of mutations in the PCFT gene, coding for an intestinal folate transporter, that are the basis for hereditary folate malabsorption.","volume":"110","author":"Zhao","year":"2007","journal-title":"Blood"},{"issue":"3","key":"2019111723170437400_B7","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1016\/j.jpeds.2008.04.009","article-title":"The clinical course and genetic defect in the PCFT in a 27-year-old woman with hereditary folate malabsorption.","volume":"153","author":"Min","year":"2008","journal-title":"J Pediatr"},{"issue":"5","key":"2019111723170437400_B8","doi-asserted-by":"crossref","first-page":"2055","DOI":"10.1182\/blood-2008-04-150276","article-title":"A novel loss of function mutation in the proton-coupled folate transporter from a patient with hereditary folate malabsorption reveals that Arg 113 is crucial for function.","volume":"112","author":"Lasry","year":"2008","journal-title":"Blood"},{"issue":"3","key":"2019111723170437400_B9","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.clim.2009.08.006","article-title":"Reversible severe combined immunodeficiency phenotype secondary to a mutation of the proton-coupled folate transporter.","volume":"133","author":"Borzutzky","year":"2009","journal-title":"Clin Immunol"},{"issue":"8","key":"2019111723170437400_B10","doi-asserted-by":"crossref","first-page":"614","DOI":"10.3109\/08880018.2010.481705","article-title":"Mutation of the proton-coupled folate transporter gene (PCFT-SLC46A1) in Turkish siblings with hereditary folate malabsorption.","volume":"27","author":"Atabay","year":"2010","journal-title":"Pediatr Hematol Oncol"},{"issue":"3","key":"2019111723170437400_B11","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/j.ymgme.2009.11.004","article-title":"A novel PCFT gene mutation (p.Cys66LeufsX99) causing hereditary folate malabsorption.","volume":"99","author":"Meyer","year":"2010","journal-title":"Mol Genet Metab"},{"issue":"5","key":"2019111723170437400_B12","doi-asserted-by":"crossref","first-page":"C1153","DOI":"10.1152\/ajpcell.00113.2010","article-title":"Properties of the Arg376 residue of the proton-coupled folate transporter (PCFT-SLC46A1) and a glutamine mutant causing hereditary folate malabsorption.","volume":"299","author":"Mahadeo","year":"2010","journal-title":"Am J Physiol Cell Physiol"},{"issue":"11","key":"2019111723170437400_B13","doi-asserted-by":"crossref","first-page":"5299","DOI":"10.1167\/iovs.07-0288","article-title":"Cloning and functional characterization of the proton-coupled electrogenic folate transporter and analysis of its expression in retinal cell types.","volume":"48","author":"Umapathy","year":"2007","journal-title":"Invest Ophthalmol Vis Sci"},{"issue":"3","key":"2019111723170437400_B14","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1124\/mol.108.045443","article-title":"The proton-coupled folate transporter (PCFT): impact on pemetrexed transport and on antifolate activities as compared to the reduced folate carrier.","volume":"74","author":"Zhao","year":"2008","journal-title":"Mol Pharmacol"},{"issue":"5","key":"2019111723170437400_B15","doi-asserted-by":"crossref","first-page":"C1669","DOI":"10.1152\/ajpcell.00202.2007","article-title":"Rodent intestinal folate transporters (SLC46A1): secondary structure, functional properties, and response to dietary folate restriction.","volume":"293","author":"Qiu","year":"2007","journal-title":"Am J Physiol Cell Physiol"},{"issue":"1","key":"2019111723170437400_B16","doi-asserted-by":"crossref","first-page":"C66","DOI":"10.1152\/ajpcell.00096.2009","article-title":"Role of the glutamate 185 residue in proton translocation mediated by the proton-coupled folate transporter SLC46A1.","volume":"297","author":"Unal","year":"2009","journal-title":"Am J Physiol Cell Physiol"},{"issue":"26","key":"2019111723170437400_B17","doi-asserted-by":"crossref","first-page":"17846","DOI":"10.1074\/jbc.M109.008060","article-title":"The functional roles of the His247 and His281 residues in folate and proton translocation mediated by the human proton-coupled folate transporter, SLC46A1.","volume":"284","author":"Unal","year":"2009","journal-title":"J Biol Chem"},{"issue":"13","key":"2019111723170437400_B18","doi-asserted-by":"crossref","first-page":"2925","DOI":"10.1021\/bi9021439","article-title":"Membrane topologicical analysis of the proton-coupled folate transporter (PCFT-SLC46A1) by the substituted cysteine accessibility method.","volume":"49","author":"Zhao","year":"2010","journal-title":"Biochemistry"},{"issue":"3","key":"2019111723170437400_B19","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1016\/j.bbrc.2009.06.007","article-title":"Hereditary folate malabsorption: a positively charged amino acid at position 113 of the proton-coupled folate transporter (PCFT\/SLC46A1) is required for folic acid binding.","volume":"386","author":"Lasry","year":"2009","journal-title":"Biochem Biophys Res Commun"},{"issue":"9","key":"2019111723170437400_B20","first-page":"3687","article-title":"The mechanism of transport of the multitargeted antifolate, MTA-LY231514, and its cross-resistance pattern in cell with impaired transport of methotrexate.","volume":"6","author":"Zhao","year":"2000","journal-title":"Clin Cancer Res"},{"issue":"6","key":"2019111723170437400_B21","doi-asserted-by":"crossref","first-page":"1407","DOI":"10.1016\/j.bbamem.2008.03.009","article-title":"N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT).","volume":"1178","author":"Unal","year":"2008","journal-title":"Biochim Biophys Acta"},{"issue":"2","key":"2019111723170437400_B22","doi-asserted-by":"crossref","first-page":"718","DOI":"10.1158\/1078-0432.CCR-1066-03","article-title":"A prominent low-pH methotrexate transport activity in human solid tumor cells: contribution to the preservation of methotrexate pharmacological activity in HeLa cells lacking the reduced folate carrier.","volume":"10","author":"Zhao","year":"2004","journal-title":"Clin Cancer Res"},{"issue":"8","key":"2019111723170437400_B23","doi-asserted-by":"crossref","first-page":"2424","DOI":"10.1158\/1535-7163.MCT-08-0938","article-title":"Hypermethylation of the human proton-coupled folate transporter (SLC46A1) minimal transcriptional regulatory region in an antifolate-resistant HeLa cell line.","volume":"8","author":"Diop-Bove","year":"2009","journal-title":"Mol Cancer Ther"},{"issue":"5","key":"2019111723170437400_B24","doi-asserted-by":"crossref","first-page":"926","DOI":"10.2144\/00285st06","article-title":"Rapid determination of membrane transport parameters in adherent cells.","volume":"28","author":"Sharif","year":"2000","journal-title":"BioTechniques"},{"issue":"1","key":"2019111723170437400_B25","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.bcp.2009.07.019","article-title":"Structure-function analysis of the highly conserved charged residues of the membrane protein FT1, the main folic acid transporter of the protozoan parasite, Leishmania.","volume":"79","author":"Dridi","year":"2010","journal-title":"Biochem Pharmacol"},{"issue":"1","key":"2019111723170437400_B26","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/0022-2836(84)90446-7","article-title":"Intrahelical hydrogen bonding of serine, threonine, and cysteine residues within alpha-helices and its relevance to membrane-bound proteins.","volume":"175","author":"Gray","year":"1984","journal-title":"J Mol Biol"},{"issue":"1","key":"2019111723170437400_B27","doi-asserted-by":"crossref","first-page":"C233","DOI":"10.1152\/ajpcell.00468.2007","article-title":"Apical membrane targeting and trafficking of the human proton-coupled transporter in polarized epithelia.","volume":"294","author":"Subramanian","year":"2008","journal-title":"Am J Physiol Cell Physiol"},{"issue":"2-3","key":"2019111723170437400_B28","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/S0014-5793(96)01181-7","article-title":"Conservation of amino acids in multiple alignments: aspartic acid has unexpected conservation.","volume":"397","author":"Fiser","year":"1996","journal-title":"FEBS Lett"},{"key":"2019111723170437400_B29","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1146\/annurev.biophys.35.040405.102005","article-title":"Lessons from lactose permease.","volume":"35","author":"Guan","year":"2006","journal-title":"Annu Rev Biophys Biomol Struct"},{"issue":"4","key":"2019111723170437400_B30","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1016\/j.jmb.2008.04.015","article-title":"The Cys154\u2013>Gly mutation in LacY causes constitutive opening of the hydrophilic periplasmic pathway.","volume":"379","author":"Nie","year":"2008","journal-title":"J Mol Biol"},{"issue":"1","key":"2019111723170437400_B31","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1097\/00005792-200201000-00004","article-title":"Hereditary folate malabsorption: family report and review of the literature.","volume":"81","author":"Geller","year":"2002","journal-title":"Medicine (Baltimore)"},{"key":"2019111723170437400_B32","article-title":"GeneReviews [Internet].","author":"Mahadeo","year":"1993"},{"issue":"7","key":"2019111723170437400_B33","doi-asserted-by":"crossref","first-page":"4267","DOI":"10.1074\/jbc.M807665200","article-title":"A role for the proton-coupled folate transporter (PCFT; SLC46A1) in folate receptor-mediated endocytosis.","volume":"284","author":"Zhao","year":"2009","journal-title":"J Biol Chem"},{"issue":"3","key":"2019111723170437400_B34","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/j.ymgme.2008.07.004","article-title":"Technical and biochemical factors affecting cerebrospinal fluid 5-MTHF, biopterin and neopterin concentrations.","volume":"95","author":"Verbeek","year":"2008","journal-title":"Mol Genet Metab"},{"issue":"1","key":"2019111723170437400_B35","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0005-2736(01)00340-6","article-title":"Localization of the murine reduced folate carrier as assessed by immunohistochemical analysis.","volume":"1513","author":"Wang","year":"2001","journal-title":"Biochim Biophys Acta"},{"issue":"3","key":"2019111723170437400_B36","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1016\/j.ajhg.2009.08.005","article-title":"Folate receptor alpha defect causes cerebral folate transport deficiency: a treatable neurodegenerative disorder associated with disturbed myelin metabolism.","volume":"85","author":"Steinfeld","year":"2009","journal-title":"Am J Hum Genet"},{"issue":"24","key":"2019111723170437400_B37","doi-asserted-by":"crossref","first-page":"2127","DOI":"10.1212\/WNL.0b013e3181c679df","article-title":"Congenital null mutations of the FOLR1 gene: a progressive neurologic disease and its treatment.","volume":"73","author":"Cario","year":"2009","journal-title":"Neurology"}],"container-title":["Blood"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/ashpublications.org\/blood\/article-pdf\/116\/24\/5162\/1461667\/zh805010005162.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/ashpublications.org\/blood\/article-pdf\/116\/24\/5162\/1461667\/zh805010005162.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2019,11,18]],"date-time":"2019-11-18T04:17:28Z","timestamp":1574050648000},"score":1,"resource":{"primary":{"URL":"https:\/\/ashpublications.org\/blood\/article\/116\/24\/5162\/28043\/Functional-roles-of-aspartate-residues-of-the"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2010,12,9]]},"references-count":37,"journal-issue":{"issue":"24","published-print":{"date-parts":[[2010,12,9]]}},"URL":"https:\/\/doi.org\/10.1182\/blood-2010-06-291237","relation":{},"ISSN":["0006-4971","1528-0020"],"issn-type":[{"value":"0006-4971","type":"print"},{"value":"1528-0020","type":"electronic"}],"subject":[],"published":{"date-parts":[[2010,12,9]]}}}