Thymidine transport in cultured mammalian cells. Kinetic analysis, temperature dependence and specificity of the transport system

Wohlhueter, Robert M.; März, Richard; Plagemann, Peter G.W.

doi:10.1016/0005-2736(79)90231-1

Cited by 90 publications

(32 citation statements)

References 24 publications

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“…In contrast, both Walker 256 and Novikoff UA cells were relatively insensitive to NBMPR with 50% inhibition of transport resulting from exposure to NBMPR concentrations greater than 10 gM. These high IC50 values are in agreement with previous reports (Wohlhueter et al, 1979;Paterson et al, 1983a;Belt, 1983) NBMPR. The concentration-effect curve for NBMPR inhibition of adenosine transport in L1210 cells was biphasic with a broad plateau spanning the range from 10 to 1000 nM-NBMPR, indicating that these cells have two types of transport activity that differ in sensitivity to NBMPR (Belt, 1983 Interaction of these compounds with the nucleoside transport mechanism ofNovikoff cells has been described (Plagemann & Wohlhueter, 1980).…”

Section: Inhibition Of Qdenosine Transport By Nbmprsupporting

confidence: 90%

Photoaffinity labelling of a nitrobenzylthioinosine-binding polypeptide from cultured Novikoff hepatoma cells

Gati¹,

Belt²,

Jakobs³

et al. 1986

Biochemical Journal

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Site-specific binding of nitrobenzylthioinosine (NBMPR) to plasma membranes of some animal cells results in the inhibition of the facilitated diffusion of nucleosides. The present study showed that nucleoside transport in Novikoff UA rat hepatoma cells is insensitive to site-saturating concentrations of NBMPR. Equilibrium binding experiments demonstrated the presence of high-affinity sites for NBMPR in a membrane-enriched fraction from these cells. In the presence of uridine or dipyridamole, specific binding of NBMPR at these sites was inhibited. When Novikoff UA membranes were covalently labelled with [3H]NBMPR by using photoaffinity techniques, specifically bound radioactivity was incorporated exclusively into a polypeptide(s) with an apparent Mr of 72,000-80,000, determined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Covalent labelling of this polypeptide was abolished in the presence of excess nitrobenzylthioguanosine (NBTGR) and reduced in the presence of adenosine, uridine or dipyridamole. The apparent Mr of the NBMPR-binding polypeptide in Novikoff UA cells is significantly higher than that reported for corresponding polypeptides in other cell types (Mr 45,000-66,000). When membrane-enriched preparations from S49 mouse lymphoma cells were photolabelled and mixed with labelled NovikoffUA membrane-enriched preparations, gel electrophoresis resolved the NBMPR-binding polypeptides from the two preparations.

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Section: Inhibition Of Qdenosine Transport By Nbmprsupporting

confidence: 90%

Photoaffinity labelling of a nitrobenzylthioinosine-binding polypeptide from cultured Novikoff hepatoma cells

Gati¹,

Belt²,

Jakobs³

et al. 1986

Biochemical Journal

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“…Exogenous purine and pyrimidine nucleosides, including thymidine, are rapidly incorporated via a variety of salvage pathways (27). Incorporation is a complex process involving transport of the nucleosides across the cell membrane (13,14), multiple phosphorylations resulting in their entry into cellular nucleotide pools, and final polymerization into nucleic acids for proliferation, protein synthesis (15), and DNA maintenance and repair (16).…”

Section: Discussionmentioning

confidence: 99%

Hydrogen Peroxide-Induced Thymidine Incorporation Into Cultured Rat Astrocytes

2006

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“…Increased 3-O-methylglucose uptake is regularly seen upon incubation with inhibitors of energy metabolism [28,29]. Plagemann and co-workers using Novikoff rat hepatoma cells demonstrated that ATP depletion did not affect transport of TdR or Urd, but effectively reduced phosphorylation of these nucleosides in situ, resulting in a decrease in macromolecular synthesis [36][37][38].…”

Section: Discussionmentioning

confidence: 99%

Effects of tri-n-butyltin chloride on energy metabolism, macromolecular synthesis, precursor uptake and cyclic AMP production in isolated rat thymocytes

Snoeij

Punt

Penninks

et al. 1986

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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The inhibitor of oxidative phosphorylation tri-n-butyltin chloride (TBTC) causes membrane damage and disintegration of isolated rat thymocytes at concentrations higher than 1 ixM. From a concentration of 0.1 I~M, TBTC disturbs energy metabolism as indicated by an increase in methylglucose uptake, glucose consumption and lactate production and by a decrease in cellular ATP levels. Over the same TBTC concentration range, the incorporation of DNA, RNA and protein precursors are markedly reduced. Moreover the production of cyclic AMP upon stimulation of the cells with prostaglandin E 1 is effectively inhibited. These effects cannot be explained by an inhibition of nucleoside kinase activity, amino acid uptake or adenylate cyclase activity. The effects of TBTC on macromolecular synthesis and cyclic AMP production are possibly due to a disturbance of the cellular energy state.

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Thymidine transport in cultured mammalian cells. Kinetic analysis, temperature dependence and specificity of the transport system

Cited by 90 publications

References 24 publications

Photoaffinity labelling of a nitrobenzylthioinosine-binding polypeptide from cultured Novikoff hepatoma cells

Photoaffinity labelling of a nitrobenzylthioinosine-binding polypeptide from cultured Novikoff hepatoma cells

Hydrogen Peroxide-Induced Thymidine Incorporation Into Cultured Rat Astrocytes

Effects of tri-n-butyltin chloride on energy metabolism, macromolecular synthesis, precursor uptake and cyclic AMP production in isolated rat thymocytes

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