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. 1995 Jun 15;308(Pt 3):725–731. doi: 10.1042/bj3080725

Kinetics of extracellular ATP hydrolysis by microvascular endothelial cells from rat heart.

P Meghji 1, J D Pearson 1, L L Slakey 1
PMCID: PMC1136785  PMID: 8948425

Abstract

We have characterized the ectonucleotidases that catalyse the reaction sequence ATP-->ADP-->AMP-->adenosine on microvascular endothelial cells cultured from the rat heart. Computer simulation and data fitting of progress of reaction curves showed that depletion of substrate at the cell surface dominates the regulation of the rate of hydrolysis of ATP when it is presented to the cells. Preferential delivery of AMP by ADPase to 5'-nucleotidase makes a significant contribution to the regulation of adenosine production from ATP or ADP. By contrast, we found no evidence for the preferential delivery of ADP from ATPase to ADPase. Feed-forward inhibition of AMP hydrolysis by ADP and/or ATP also modulated the rate of adenosine production. The properties of the ectonucleotidases on rat heart microvascular cells are such that adenosine is produced at a steady rate over a wide range of ATP concentrations.

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Selected References

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  1. Ager A., Gordon J. L., Moncada S., Pearson J. D., Salmon J. A., Trevethick M. A. Effects of isolation and culture on prostaglandin synthesis by porcine aortic endothelial and smooth muscle cells. J Cell Physiol. 1982 Jan;110(1):9–16. doi: 10.1002/jcp.1041100103. [DOI] [PubMed] [Google Scholar]
  2. Alroy J., Goyal V., Skutelsky E. Lectin histochemistry of mammalian endothelium. Histochemistry. 1987;86(6):603–607. doi: 10.1007/BF00489554. [DOI] [PubMed] [Google Scholar]
  3. Anderson R. G. Plasmalemmal caveolae and GPI-anchored membrane proteins. Curr Opin Cell Biol. 1993 Aug;5(4):647–652. doi: 10.1016/0955-0674(93)90135-d. [DOI] [PubMed] [Google Scholar]
  4. Bruns R. F. Adenosine receptors. Roles and pharmacology. Ann N Y Acad Sci. 1990;603:211–226. doi: 10.1111/j.1749-6632.1990.tb37674.x. [DOI] [PubMed] [Google Scholar]
  5. Burger R. M., Lowenstein J. M. 5'-Nucleotidase from smooth muscle of small intestine and from brain. Inhibition of nucleotides. Biochemistry. 1975 Jun 3;14(11):2362–2366. doi: 10.1021/bi00682a014. [DOI] [PubMed] [Google Scholar]
  6. Coade S. B., Pearson J. D. Metabolism of adenine nucleotides in human blood. Circ Res. 1989 Sep;65(3):531–537. doi: 10.1161/01.res.65.3.531. [DOI] [PubMed] [Google Scholar]
  7. Cusack N. J., Pearson J. D., Gordon J. L. Stereoselectivity of ectonucleotidases on vascular endothelial cells. Biochem J. 1983 Sep 15;214(3):975–981. doi: 10.1042/bj2140975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Diglio C. A., Grammas P., Giacomelli F., Wiener J. Rat heart-derived endothelial and smooth muscle cell cultures: isolation, cloning and characterization. Tissue Cell. 1988;20(4):477–492. doi: 10.1016/0040-8166(88)90051-1. [DOI] [PubMed] [Google Scholar]
  9. Gordon E. L., Pearson J. D., Dickinson E. S., Moreau D., Slakey L. L. The hydrolysis of extracellular adenine nucleotides by arterial smooth muscle cells. Regulation of adenosine production at the cell surface. J Biol Chem. 1989 Nov 15;264(32):18986–18995. [PubMed] [Google Scholar]
  10. Gordon E. L., Pearson J. D., Dickinson E. S., Moreau D., Slakey L. L. The hydrolysis of extracellular adenine nucleotides by arterial smooth muscle cells. Regulation of adenosine production at the cell surface. J Biol Chem. 1989 Nov 15;264(32):18986–18995. [PubMed] [Google Scholar]
  11. Gordon E. L., Pearson J. D., Slakey L. L. The hydrolysis of extracellular adenine nucleotides by cultured endothelial cells from pig aorta. Feed-forward inhibition of adenosine production at the cell surface. J Biol Chem. 1986 Nov 25;261(33):15496–15507. [PubMed] [Google Scholar]
  12. Imai S., Chin W. P., Jin H., Nakazawa M. Production of AMP and adenosine in the interstitial fluid compartment of the isolated perfused normoxic guinea pig heart. Pflugers Arch. 1989 Aug;414(4):443–449. doi: 10.1007/BF00585055. [DOI] [PubMed] [Google Scholar]
  13. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lin S. H., Russell W. E. Two Ca2+-dependent ATPases in rat liver plasma membrane. The previously purified (Ca2+-Mg2+)-ATPase is not a Ca2+-pump but an ecto-ATPase. J Biol Chem. 1988 Sep 5;263(25):12253–12258. [PubMed] [Google Scholar]
  15. Lin S. H. The rat liver plasma membrane high affinity (Ca2+-Mg2+)-ATPase is not a calcium pump. Comparison with ATP-dependent calcium transporter. J Biol Chem. 1985 Sep 15;260(20):10976–10980. [PubMed] [Google Scholar]
  16. Meghji P., Holmquist C. A., Newby A. C. Adenosine formation and release from neonatal-rat heart cells in culture. Biochem J. 1985 Aug 1;229(3):799–805. doi: 10.1042/bj2290799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Meghji P., Pearson J. D., Slakey L. L. Regulation of extracellular adenosine production by ectonucleotidases of adult rat ventricular myocytes. Am J Physiol. 1992 Jul;263(1 Pt 2):H40–H47. doi: 10.1152/ajpheart.1992.263.1.H40. [DOI] [PubMed] [Google Scholar]
  18. Meghji P., Skladanowski A. C., Newby A. C., Slakey L. L., Pearson J. D. Effect of 5'-deoxy-5'-isobutylthioadenosine on formation and release of adenosine from neonatal and adult rat ventricular myocytes. Biochem J. 1993 May 1;291(Pt 3):833–839. doi: 10.1042/bj2910833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Naito Y., Lowenstein J. M. 5'-Nucleotidase from rat heart. Biochemistry. 1981 Sep 1;20(18):5188–5194. doi: 10.1021/bi00521a014. [DOI] [PubMed] [Google Scholar]
  20. Needham L., Cusack N. J., Pearson J. D., Gordon J. L. Characteristics of the P2 purinoceptor that mediates prostacyclin production by pig aortic endothelial cells. Eur J Pharmacol. 1987 Feb 10;134(2):199–209. doi: 10.1016/0014-2999(87)90166-x. [DOI] [PubMed] [Google Scholar]
  21. Nishida M., Carley W. W., Gerritsen M. E., Ellingsen O., Kelly R. A., Smith T. W. Isolation and characterization of human and rat cardiac microvascular endothelial cells. Am J Physiol. 1993 Feb;264(2 Pt 2):H639–H652. doi: 10.1152/ajpheart.1993.264.2.H639. [DOI] [PubMed] [Google Scholar]
  22. Norman G. A., Follett M. J., Hector D. A. Quantitative thin-layer chromatography of ATP and the products of its degradation in meat tissue. J Chromatogr. 1974 Mar 13;90(1):105–111. doi: 10.1016/s0021-9673(01)94779-x. [DOI] [PubMed] [Google Scholar]
  23. Olsson R. A., Pearson J. D. Cardiovascular purinoceptors. Physiol Rev. 1990 Jul;70(3):761–845. doi: 10.1152/physrev.1990.70.3.761. [DOI] [PubMed] [Google Scholar]
  24. Paddle B. M., Burnstock G. Release of ATP from perfused heart during coronary vasodilatation. Blood Vessels. 1974;11(3):110–119. doi: 10.1159/000158005. [DOI] [PubMed] [Google Scholar]
  25. Pearson J. D., Carleton J. S., Gordon J. L. Metabolism of adenine nucleotides by ectoenzymes of vascular endothelial and smooth-muscle cells in culture. Biochem J. 1980 Aug 15;190(2):421–429. doi: 10.1042/bj1900421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pearson J. D., Coade S. B., Cusack N. J. Characterization of ectonucleotidases on vascular smooth-muscle cells. Biochem J. 1985 Sep 1;230(2):503–507. doi: 10.1042/bj2300503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pronk A., Leguit P., Hoynck van Papendrecht A. A., Hagelen E., van Vroonhoven T. J., Verbrugh H. A. A cobblestone cell isolated from the human omentum: the mesothelial cell; isolation, identification, and growth characteristics. In Vitro Cell Dev Biol. 1993 Feb;29A(2):127–134. doi: 10.1007/BF02630943. [DOI] [PubMed] [Google Scholar]
  28. Ragazzi E., Wu S. N., Shryock J., Belardinelli L. Electrophysiological and receptor binding studies to assess activation of the cardiac adenosine receptor by adenine nucleotides. Circ Res. 1991 Apr;68(4):1035–1044. doi: 10.1161/01.res.68.4.1035. [DOI] [PubMed] [Google Scholar]
  29. Ronca-Testoni S., Borghini F. Degradation of perfused adenine compounds up to uric acid in isolated rat heart. J Mol Cell Cardiol. 1982 Mar;14(3):177–180. doi: 10.1016/0022-2828(82)90116-x. [DOI] [PubMed] [Google Scholar]
  30. Slakey L. L., Cosimini K., Earls J. P., ThomasC, Gordon E. L. Simulation of extracellular nucleotide hydrolysis and determination of kinetic constants for the ectonucleotidases. J Biol Chem. 1986 Nov 25;261(33):15505–15507. [PubMed] [Google Scholar]
  31. Yagi K., Shinbo M., Hashizume M., Shimba L. S., Kurimura S., Miura Y. ATP diphosphohydrolase is responsible for ecto-ATPase and ecto-ADPase activities in bovine aorta endothelial and smooth muscle cells. Biochem Biophys Res Commun. 1991 Nov 14;180(3):1200–1206. doi: 10.1016/s0006-291x(05)81323-3. [DOI] [PubMed] [Google Scholar]
  32. van Hinsbergh V. W., Kooistra T., Scheffer M. A., Hajo van Bockel J., van Muijen G. N. Characterization and fibrinolytic properties of human omental tissue mesothelial cells. Comparison with endothelial cells. Blood. 1990 Apr 1;75(7):1490–1497. [PubMed] [Google Scholar]

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