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Article: Genomic organization and functional characterization of the human concentrative nucleoside transporter-3 isoform (hCNT3) expressed in mammalian cells

TitleGenomic organization and functional characterization of the human concentrative nucleoside transporter-3 isoform (hCNT3) expressed in mammalian cells
Authors
KeywordsAdenosine
Cos7L cells
Genome
hCNT3
Nucleoside transport
PK15NTD cells
Issue Date2003
PublisherSpringer. The Journal's web site is located at http://link.springer.de/link/service/journals/00424/index.htm
Citation
Pflugers Archiv European Journal Of Physiology, 2003, v. 447 n. 2, p. 195-204 How to Cite?
AbstractHuman CNT3 encodes the concentrative nucleoside transport N3 system. Previous expression studies in oocytes showed that the K mvalues for nucleosides of the cloned hCNT3 were 7- to 25-fold lower than the endogenous N3 transporter in HL60 cells. Therefore, in the present study we re-examined the kinetic properties of the cloned hCNT3 using mammalian cell expression systems by transient expression in Cos7L cells and stably expression in nucleoside transporter deficient PK15NTD cells. We demonstrated that hCNT3 is a Na-dependent, broadly-selective nucleoside transporter with affinities (<11 μM) for nucleosides closely resembling the endogenous N3 transporter. Pharmacological studies showed that phloridzin is a mixed-type inhibitor of hCNT3 (K i=15 μM), and the dideoxyuridine analogs are poor substrates. By epitope-tagging, we further demonstrated that hCNT3 is N-glycosylated as PNGase F and Endo H deglycosylated hCNT3 from 67 kDa to 58 kDa. Searching the human genome database, we identified the genomic organization of hCNT3. This gene contains 19 exons and its exon-intron boundaries within the coding sequence exactly match with those of hCNT1 and hCNT2 with one additional exon in the N-terminus. Our data suggest that hCNT3 gene is evolutionarily conserved with hCNT1 and hCNT2. Physiologically, hCNT3 is a glycoprotein, which transports purine and pyrimidine nucleosides in a Na-dependent manner with high affinities.
Persistent Identifierhttp://hdl.handle.net/10722/91729
ISSN
2015 Impact Factor: 3.654
2015 SCImago Journal Rankings: 1.638
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorToan, SVen_HK
dc.contributor.authorTo, KKWen_HK
dc.contributor.authorLeung, GPHen_HK
dc.contributor.authorDe Souza, MOen_HK
dc.contributor.authorWard, JLen_HK
dc.contributor.authorTse, CMen_HK
dc.date.accessioned2010-09-17T10:24:37Z-
dc.date.available2010-09-17T10:24:37Z-
dc.date.issued2003en_HK
dc.identifier.citationPflugers Archiv European Journal Of Physiology, 2003, v. 447 n. 2, p. 195-204en_HK
dc.identifier.issn0031-6768en_HK
dc.identifier.urihttp://hdl.handle.net/10722/91729-
dc.description.abstractHuman CNT3 encodes the concentrative nucleoside transport N3 system. Previous expression studies in oocytes showed that the K mvalues for nucleosides of the cloned hCNT3 were 7- to 25-fold lower than the endogenous N3 transporter in HL60 cells. Therefore, in the present study we re-examined the kinetic properties of the cloned hCNT3 using mammalian cell expression systems by transient expression in Cos7L cells and stably expression in nucleoside transporter deficient PK15NTD cells. We demonstrated that hCNT3 is a Na-dependent, broadly-selective nucleoside transporter with affinities (<11 μM) for nucleosides closely resembling the endogenous N3 transporter. Pharmacological studies showed that phloridzin is a mixed-type inhibitor of hCNT3 (K i=15 μM), and the dideoxyuridine analogs are poor substrates. By epitope-tagging, we further demonstrated that hCNT3 is N-glycosylated as PNGase F and Endo H deglycosylated hCNT3 from 67 kDa to 58 kDa. Searching the human genome database, we identified the genomic organization of hCNT3. This gene contains 19 exons and its exon-intron boundaries within the coding sequence exactly match with those of hCNT1 and hCNT2 with one additional exon in the N-terminus. Our data suggest that hCNT3 gene is evolutionarily conserved with hCNT1 and hCNT2. Physiologically, hCNT3 is a glycoprotein, which transports purine and pyrimidine nucleosides in a Na-dependent manner with high affinities.en_HK
dc.languageengen_HK
dc.publisherSpringer. The Journal's web site is located at http://link.springer.de/link/service/journals/00424/index.htmen_HK
dc.relation.ispartofPflugers Archiv European Journal of Physiologyen_HK
dc.subjectAdenosineen_HK
dc.subjectCos7L cellsen_HK
dc.subjectGenomeen_HK
dc.subjecthCNT3en_HK
dc.subjectNucleoside transporten_HK
dc.subjectPK15NTD cellsen_HK
dc.subject.meshAnimalsen_HK
dc.subject.meshCOS Cellsen_HK
dc.subject.meshCations - metabolismen_HK
dc.subject.meshCell Lineen_HK
dc.subject.meshCercopithecus aethiopsen_HK
dc.subject.meshEvolution, Molecularen_HK
dc.subject.meshGenome, Humanen_HK
dc.subject.meshGlycosylationen_HK
dc.subject.meshHumansen_HK
dc.subject.meshMembrane Transport Modulatorsen_HK
dc.subject.meshMembrane Transport Proteins - antagonists & inhibitors - genetics - physiologyen_HK
dc.subject.meshNucleoside Transport Proteins - metabolismen_HK
dc.subject.meshPhlorhizin - pharmacologyen_HK
dc.subject.meshPurine Nucleosides - metabolismen_HK
dc.subject.meshPyrimidine Nucleosides - metabolismen_HK
dc.subject.meshSodium - metabolismen_HK
dc.subject.meshTime Factorsen_HK
dc.subject.meshTransfectionen_HK
dc.subject.meshUridine - pharmacologyen_HK
dc.titleGenomic organization and functional characterization of the human concentrative nucleoside transporter-3 isoform (hCNT3) expressed in mammalian cellsen_HK
dc.typeArticleen_HK
dc.identifier.emailLeung, GPH: gphleung@hkucc.hku.hken_HK
dc.identifier.authorityLeung, GPH=rp00234en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1007/s00424-003-1166-0en_HK
dc.identifier.pmid14504928-
dc.identifier.scopuseid_2-s2.0-0842287376en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0842287376&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume447en_HK
dc.identifier.issue2en_HK
dc.identifier.spage195en_HK
dc.identifier.epage204en_HK
dc.identifier.isiWOS:000186589800010-
dc.publisher.placeGermanyen_HK
dc.identifier.scopusauthoridToan, SV=6505873802en_HK
dc.identifier.scopusauthoridTo, KKW=55316299500en_HK
dc.identifier.scopusauthoridLeung, GPH=35963668200en_HK
dc.identifier.scopusauthoridDe Souza, MO=7202375834en_HK
dc.identifier.scopusauthoridWard, JL=7404119073en_HK
dc.identifier.scopusauthoridTse, CM=7103295076en_HK

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