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DC Field | Value | Language |
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dc.contributor.author | Wiley, J S | en |
dc.contributor.author | Brocklebank, A M | en |
dc.contributor.author | Snook, M B | en |
dc.contributor.author | Jamieson, Gary P | en |
dc.contributor.author | Sawyer, W H | en |
dc.contributor.author | Craik, J D | en |
dc.contributor.author | Cass, C E | en |
dc.contributor.author | Robins, M J | en |
dc.contributor.author | McAdam, D P | en |
dc.contributor.author | Paterson, A R | en |
dc.date.accessioned | 2015-05-16T00:30:28Z | |
dc.date.available | 2015-05-16T00:30:28Z | |
dc.date.issued | 1991-02-01 | en |
dc.identifier.citation | The Biochemical Journal; 273 ( Pt 3)(): 667-72 | en |
dc.identifier.govdoc | 1996965 | en |
dc.identifier.other | PUBMED | en |
dc.identifier.uri | https://ahro.austin.org.au/austinjspui/handle/1/10930 | en |
dc.description.abstract | The 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.iso | en | en |
dc.subject.other | Adenosine.analogs & derivatives.chemical synthesis.pharmacology | en |
dc.subject.other | Affinity Labels | en |
dc.subject.other | Carrier Proteins.antagonists & inhibitors | en |
dc.subject.other | Cell Line | en |
dc.subject.other | Cytarabine.metabolism | en |
dc.subject.other | Flow Cytometry | en |
dc.subject.other | Fluorescent Dyes.chemical synthesis | en |
dc.subject.other | Humans | en |
dc.subject.other | Kinetics | en |
dc.subject.other | Membrane Proteins.antagonists & inhibitors | en |
dc.subject.other | Molecular Structure | en |
dc.subject.other | Nucleoside Transport Proteins | en |
dc.subject.other | Spectrometry, Fluorescence | en |
dc.subject.other | Thioinosine.analogs & derivatives.metabolism | en |
dc.subject.other | Thionucleosides.chemical synthesis.pharmacology | en |
dc.title | A 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.type | Journal Article | en |
dc.identifier.journaltitle | The Biochemical journal | en |
dc.identifier.affiliation | Department of Haematology, Austin Hospital, Heidelberg, Melbourne, Victoria, Australia | en |
dc.description.pages | 667-72 | en |
dc.relation.url | https://pubmed.ncbi.nlm.nih.gov/1996965 | en |
dc.type.austin | Journal Article | en |
item.openairecristype | http://purl.org/coar/resource_type/c_18cf | - |
item.cerifentitytype | Publications | - |
item.fulltext | With Fulltext | - |
item.grantfulltext | open | - |
item.languageiso639-1 | en | - |
item.openairetype | Journal Article | - |
Appears in Collections: | Journal articles |
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1996965.pdf | 1.06 MB | Adobe PDF | View/Open |
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