Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1985 Dec 15;232(3):681–688. doi: 10.1042/bj2320681

Nucleoside transport in Walker 256 rat carcinosarcoma and S49 mouse lymphoma cells. Differences in sensitivity to nitrobenzylthioinosine and thiol reagents.

J A Belt, L D Noel
PMCID: PMC1152939  PMID: 3004414

Abstract

The characteristics of nucleoside transport were examined in Walker 256 rat carcinosarcoma and S49 mouse lymphoma cells. In Walker 256 cells the initial rates of uridine, thymidine and adenosine uptake were insensitive to the nucleoside transport inhibitor nitrobenzylthioinosine (NBMPR) (1 microM), but were partially inhibited by dipyridamole (10 microM), another inhibitor of nucleoside transport. In contrast, the transport of these nucleosides in S49 cells was completely blocked by both inhibitors. Nucleoside transport in Walker 256 and S49 cells also differed in its sensitivity to the thiol reagent p-chloromercuribenzenesulphonate (pCMBS). Uridine transport in Walker 256 cells was inhibited by pCMBS with an IC50 (concentration producing 50% inhibition) of less than 25 microM, and inhibition was readily reversed by beta-mercaptoethanol. In S49 cells uridine transport was only inhibited at much higher concentrations of pCMBS (IC50 approximately equal to 300 microM). In other respects nucleoside transport in Walker 256 and S49 cells were quite similar. The Km and Vmax. values for uridine transport were nearly identical, and the transporters of both cell lines appeared to accept a broad range of nucleosides as substrates. Uridine transport in Walker 256 cells was non-concentrative and did not require an energy source. These studies demonstrate that nucleoside uptake in Walker 256 cells is mediated by a facilitated-diffusion mechanism which differs markedly from that of S49 cells in its sensitivity to the transport inhibitor NBMPR and the thiol reagent pCMBS.

Full text

PDF
681

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belt J. A. Heterogeneity of nucleoside transport in mammalian cells. Two types of transport activity in L1210 and other cultured neoplastic cells. Mol Pharmacol. 1983 Nov;24(3):479–484. [PubMed] [Google Scholar]
  2. Belt J. A. Nitrobenzylthioinosine-insensitive uridine transport in human lymphoblastoid and murine leukemia cells. Biochem Biophys Res Commun. 1983 Jan 27;110(2):417–423. doi: 10.1016/0006-291x(83)91165-8. [DOI] [PubMed] [Google Scholar]
  3. Bibi O., Schwartz J., Eilam Y., Shohami E., Cabantchik Z. I. Nucleoside transport in mammalian cell membranes. IV. Organomercurials and organomercurial-mercaptonucleoside complexes as probes for nucleoside transport systems in hamster cells. J Membr Biol. 1978 Mar 10;39(2-3):159–183. doi: 10.1007/BF01870330. [DOI] [PubMed] [Google Scholar]
  4. Cass C. E., Gaudette L. A., Paterson A. R. Mediated transport of nucleosides in human erythrocytes. Specific binding of the inhibitor nitrobenzylthioinosine to nucleoside transport sites in the erythrocyte membrane. Biochim Biophys Acta. 1974 Apr 12;345(1):1–10. doi: 10.1016/0005-2736(74)90239-9. [DOI] [PubMed] [Google Scholar]
  5. Cass C. E., Kolassa N., Uehara Y., Dahlig-Harley E., Harley E. R., Paterson A. R. Absence of binding sites for the transport inhibitor nitrobenzylthioinosine on nucleoside transport-deficient mouse lymphoma cells. Biochim Biophys Acta. 1981 Dec 21;649(3):769–777. doi: 10.1016/0005-2736(81)90182-6. [DOI] [PubMed] [Google Scholar]
  6. Cleland W. W. Statistical analysis of enzyme kinetic data. Methods Enzymol. 1979;63:103–138. doi: 10.1016/0076-6879(79)63008-2. [DOI] [PubMed] [Google Scholar]
  7. Cohen A., Ullman B., Martin D. W., Jr Characterization of a mutant mouse lymphoma cell with deficient transport of purine and pyrimidine nucleosides. J Biol Chem. 1979 Jan 10;254(1):112–116. [PubMed] [Google Scholar]
  8. Cybulski R. L., Fry D. W., Goldman I. D. Approaches to the measurement of intracellular adenine and the discrimination between adenine transport and metabolism in L1210 leukemia cells. Biochim Biophys Acta. 1982 Feb 25;714(3):435–441. doi: 10.1016/0304-4165(82)90151-9. [DOI] [PubMed] [Google Scholar]
  9. Dahlig-Harley E., Eilam Y., Paterson A. R., Cass C. E. Binding of nitrobenzylthioinosine to high-affinity sites on the nucleoside-transport mechanism of HeLa cells. Biochem J. 1981 Nov 15;200(2):295–305. doi: 10.1042/bj2000295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eilam Y., Cabantchik Z. I. Nucleoside transport in mammalian cell membranes: a specific inhibitory mechanism of high affinity probes. J Cell Physiol. 1977 Aug;92(2):185–201. doi: 10.1002/jcp.1040920207. [DOI] [PubMed] [Google Scholar]
  11. Eilam Y., Carbantchik I. The mechanism of interaction between high-affinity probes and the uridine transport system of mammalian cells. J Cell Physiol. 1976 Dec;89(4):831–838. doi: 10.1002/jcp.1040890451. [DOI] [PubMed] [Google Scholar]
  12. Harley E. R., Paterson A. R., Cass C. E. Initial rate kinetics of the transport of adenosine and 4-amino-7-(beta-D-ribofuranosyl)pyrrolo[2,3-d]pyrimidine (tubercidin) in cultured cells. Cancer Res. 1982 Apr;42(4):1289–1295. [PubMed] [Google Scholar]
  13. Jarvis S. M., Young J. D. Nucleoside translocation in sheep reticulocytes and fetal erythrocytes: a proposed model for the nucleoside transporter. J Physiol. 1982 Mar;324:47–66. doi: 10.1113/jphysiol.1982.sp014100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lauzon G. J., Paterson A. R. Binding of the nucleoside transport inhibitor nitrobenzylthioinosine to HeLa cells. Mol Pharmacol. 1977 Sep;13(5):883–891. [PubMed] [Google Scholar]
  15. Paterson A. R., Kolassa N., Cass C. E. Transport of nucleoside drugs in animal cells. Pharmacol Ther. 1981;12(3):515–536. doi: 10.1016/0163-7258(81)90096-6. [DOI] [PubMed] [Google Scholar]
  16. Paterson A. R., Lau E. Y., Dahlig E., Cass C. E. A common basis for inhibition of nucleoside transport by dipyridamole and nitrobenzylthioinosine? Mol Pharmacol. 1980 Jul;18(1):40–44. [PubMed] [Google Scholar]
  17. Paul B., Chen M. F., Paterson A. R. Inhibitors of nucleoside transport. A structure-activity study using human erythrocytes. J Med Chem. 1975 Oct;18(10):968–973. doi: 10.1021/jm00244a003. [DOI] [PubMed] [Google Scholar]
  18. Plagemann P. G., Marz R., Wohlhueter R. M. Uridine transport in Novikoff rat hepatoma cells and other cell lines and its relationship to uridine phosphorylation and phosphorolysis. J Cell Physiol. 1978 Oct;97(1):49–72. doi: 10.1002/jcp.1040970107. [DOI] [PubMed] [Google Scholar]
  19. Plagemann P. G., Wohlhueter R. M. Effect of sulfhydryl reagents on nucleoside transport in cultured mammalian cells. Arch Biochem Biophys. 1984 Sep;233(2):489–500. doi: 10.1016/0003-9861(84)90472-7. [DOI] [PubMed] [Google Scholar]
  20. Plagemann P. G., Wohlhueter R. M. Nucleoside transport in cultured mammalian cells. Multiple forms with different sensitivity to inhibition by nitrobenzylthioinosine or hypoxanthine. Biochim Biophys Acta. 1984 Jun 13;773(1):39–52. doi: 10.1016/0005-2736(84)90548-0. [DOI] [PubMed] [Google Scholar]
  21. Wohlhueter R. M., Marz R., Graff J. C., Plagemann P. G. A rapid-mixing technique to measure transport in suspended animal cells: applications to nucleoside transport in Novikoff rat hepatoma cells. Methods Cell Biol. 1978;20:211–236. doi: 10.1016/s0091-679x(08)62020-8. [DOI] [PubMed] [Google Scholar]
  22. Wohlhueter R. M., Marz R., Plagemann P. G. Properties of the thymidine transport system of Chinese hamster ovary cells as probed by nitrobenzylthioinosine. J Membr Biol. 1978 Sep 19;42(3):247–264. doi: 10.1007/BF01870361. [DOI] [PubMed] [Google Scholar]
  23. Wohlhueter R. M., Marz R., Plagemann P. G. Thymidine transport in cultured mammalian cells. Kinetic analysis, temperature dependence and specificity of the transport system. Biochim Biophys Acta. 1979 May 17;553(2):262–283. doi: 10.1016/0005-2736(79)90231-1. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES