Please use this identifier to cite or link to this item: https://ahro.austin.org.au/austinjspui/handle/1/10930
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dc.contributor.authorWiley, J Sen
dc.contributor.authorBrocklebank, A Men
dc.contributor.authorSnook, M Ben
dc.contributor.authorJamieson, Gary Pen
dc.contributor.authorSawyer, W Hen
dc.contributor.authorCraik, J Den
dc.contributor.authorCass, C Een
dc.contributor.authorRobins, M Jen
dc.contributor.authorMcAdam, D Pen
dc.contributor.authorPaterson, A Ren
dc.date.accessioned2015-05-16T00:30:28Z
dc.date.available2015-05-16T00:30:28Z
dc.date.issued1991-02-01en
dc.identifier.citationThe Biochemical Journal; 273 ( Pt 3)(): 667-72en
dc.identifier.govdoc1996965en
dc.identifier.otherPUBMEDen
dc.identifier.urihttps://ahro.austin.org.au/austinjspui/handle/1/10930en
dc.description.abstractThe N6-(4-nitrobenzyl) derivative of adenosine is a tight-binding inhibitor of the equilibrative inhibitor-sensitive nucleoside transporter of mammalian cells. A fluorescent ligand for this transporter has been synthesized by allowing an adenosine analogue. 5'-S-(2-aminoethyl)-N6-(4-nitrobenzyl)-5'-thioadenosine (SAENTA), to react with fluorescein isothiocyanate. The purified adduct had a SAENTA/fluorescein molar ratio of 0.92:1 calculated from its absorption spectrum. The intensity of fluorescent emission from the SAENTA-chi 2-fluorescein adduct was 30% that of fluorescein isothiocyanate (chi 2 is the number of atoms in the linkage between fluorescein and SAENTA). SAENTA-chi 2-fluorescein inhibited the influx of nucleosides into cultured leukaemic cells with an IC50 (total concentration of inhibitor producing 50% inhibition) of 40 nM. The adduct inhibited the binding of [3H]nitrobenzylthioinosine ([3H]NBMPR) with half-maximal inhibition at 50-100 nM. Mass Law analysis of the competitive-binding data suggested the presence of two classes of sites for [3H]NBMPR binding, only one of which was accessible to SAENTA-chi 2-fluorescein. Flow cytometry was used to analyse equilibrium binding of SAENTA-chi 2-fluorescein to leukaemic cells and a Kd of 6 nM was obtained. SAENTA-chi 2-fluorescein is a high-affinity ligand for the equilibrative inhibitor-sensitive nucleoside transporter which allows rapid assessment of transport capacity by flow cytometry.en
dc.language.isoenen
dc.subject.otherAdenosine.analogs & derivatives.chemical synthesis.pharmacologyen
dc.subject.otherAffinity Labelsen
dc.subject.otherCarrier Proteins.antagonists & inhibitorsen
dc.subject.otherCell Lineen
dc.subject.otherCytarabine.metabolismen
dc.subject.otherFlow Cytometryen
dc.subject.otherFluorescent Dyes.chemical synthesisen
dc.subject.otherHumansen
dc.subject.otherKineticsen
dc.subject.otherMembrane Proteins.antagonists & inhibitorsen
dc.subject.otherMolecular Structureen
dc.subject.otherNucleoside Transport Proteinsen
dc.subject.otherSpectrometry, Fluorescenceen
dc.subject.otherThioinosine.analogs & derivatives.metabolismen
dc.subject.otherThionucleosides.chemical synthesis.pharmacologyen
dc.titleA new fluorescent probe for the equilibrative inhibitor-sensitive nucleoside transporter. 5'-S-(2-aminoethyl)-N6-(4-nitrobenzyl)-5'-thioadenosine (SAENTA)-chi 2-fluorescein.en
dc.typeJournal Articleen
dc.identifier.journaltitleThe Biochemical journalen
dc.identifier.affiliationDepartment of Haematology, Austin Hospital, Heidelberg, Melbourne, Victoria, Australiaen
dc.description.pages667-72en
dc.relation.urlhttps://pubmed.ncbi.nlm.nih.gov/1996965en
dc.type.austinJournal Articleen
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.cerifentitytypePublications-
item.fulltextWith Fulltext-
item.grantfulltextopen-
item.languageiso639-1en-
item.openairetypeJournal Article-
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