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Prostaglandin Endoperoxide Synthase from Human Cell Line Lu-65

  • Noreen J. Hickok
  • Mary Alosio
  • Richard S. Bookman
Part of the GWUMC Department of Biochemistry Annual Spring Symposia book series (GWUN)

Abstract

In this chapter, we present our data on the isolation and properties of prostaglandin endoperoxide synthase (PES; E.C. 1.14.99.1) from a human cell line. This enzyme has been isolated and purified to homogeneity in other species; partially purified human platelet PES that retained activity has been prepared by DEAE-chromatog-raphy (Hammarström and Falardeau, 1977) and has been purified to near-homogeneity (Ho et al., 1980). In other studies, an inactive human platelet PES has been identified by SDS-PAGE (Roth and Majerus, 1975). Preliminary data on the characterization of this PES from a human cell line, Lu-65, is presented.

Keywords

Arachidonic Acid Prostaglandin Synthetase Cyclooxygenase Activity Eicosatrienoic Acid Prostaglandin Endoperoxide 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Bockman, R.S., Bellin, A., Repo, M.A., Hickok, N.J., and Kameya, T., 1983, In vivo and in vitro biologic activities of two human cell lines derived from anaplastic lung cancers, Cancer Res. 43:4571–4576.Google Scholar
  2. Bonner, W.M., and Laskey, R.A., 1974, A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels, Eur. J. Biochem. 46:83–88.PubMedCrossRefGoogle Scholar
  3. Egan, R.W., Paxton, J., and Kuehl, F.A., Jr., 1976, Mechanism for irreversible self-deactivation of prostaglandin synthetase, J. Biol. Chem. 251:7329–7335.PubMedGoogle Scholar
  4. Egan, R.W., Gale, P.H., and Kuehl, F.A., Jr., 1979, Reduction of hydroperoxides in the prostaglandin synthetic pathway by a microsomal peroxidase, J. Biol. Chem. 254:3295–3302.PubMedGoogle Scholar
  5. Egan, R.W., Gale, P.H., Beveridge, G.C, Marnett, L.J., and Kuehl, F.A., Jr., 1980, Direct and indirect involvement of radical scavengers during prostaglandin biosynthesis, Adv. Prostaglandin Thromboxane Res. 6:153–155.PubMedGoogle Scholar
  6. Flower, R. J., and Blackwell, G. J., 1976, The importance of phospholipase A2 in prostaglandin biosynthesis, Biochem. Pharmacol. 25:285–291.PubMedCrossRefGoogle Scholar
  7. Hammarström A., and Falardeau, P., 1977, Resolution of prostaglandin endoperoxide synthase and thromboxane synthase of human platelets, Proc. Natl. Acad. Sci. U.S.A. 74:3691–3695.PubMedCrossRefGoogle Scholar
  8. Hemler, M.E., and Lands, W.E.M., 1980, Protection of cyclooxygenase activity during heme-induced destabilization, Arch. Biochem. Biophys. 201:586-593. Hemler, M. E., Lands, W. E. M., and Smith, W. L., 1976, Purification of the cyclooxygenase that forms prostaglandins. Demonstration of two forms of iron in the holoenzyme, J. Biol. Chem. 251:5575–5579.Google Scholar
  9. Ho, P.P.K., Towner, R.D., and Esterman, M.A., 1980, Purification and characterization of fatty acid cyclooxygenase from human platelets, Prep. Biochem. 10:597–613.PubMedCrossRefGoogle Scholar
  10. Kessler, S. W., 1981, Use of protein A-bearing staphylococci for the immunoprecipitation and isolation of antigens from cells, Methods Enzymol. 73:442–459.PubMedCrossRefGoogle Scholar
  11. Kulmacz, R.J., and Lands, W.E.M., 1982, Protection of prostaglandin synthase from trypsin upon binding of heme, Biochem. Biophys. Res. Commun. 104:758–764.PubMedCrossRefGoogle Scholar
  12. Laemmli, U.K., 1970, Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature 227:680–685.PubMedCrossRefGoogle Scholar
  13. Miyamoto, T., Ogino, N., Yamamoto, S., and Hayaishi, O., 1976, Purification of prostaglandin endoperoxide synthetase from bovine vesicular gland microsomes, J. Biol. Chem. 251:2629–2636.PubMedGoogle Scholar
  14. Powell, W.S., 1980, Rapid extraction of oxygenated metabolites of arachidonic acid from biological samples using octadecylsilyl silica, Prostaglandins 20:947–957.PubMedCrossRefGoogle Scholar
  15. Roth, G.J., and Majerus, P.W., 1975, The mechanism of the effect of aspirin on human platelets. I. Acetylation of a particulate fraction protein, J. Clin. Invest. 56:624–632.PubMedCrossRefGoogle Scholar
  16. Roth, G.J., Siok, C.J., and Ozols, J., 1980, Structural characteristics of prostaglandin synthetase from sheep vesicular gland, J. Biol. Chem. 255:1301–1304.PubMedGoogle Scholar
  17. Roth, G.J., Machuga, E.T., and Ozols, J., 1983, Isolation and covalent structure of the aspirin-modified, active-site region of prostaglandin synthetase, Biochemistry 22:4672–4675.PubMedCrossRefGoogle Scholar
  18. Samuelsson, B., 1965, On the incorporation of oxygen in the conversion of 8,11,14-eicosatrienoic acid to prostaglandin E1, J. Am. Cancer Soc. 87:3011–3013.CrossRefGoogle Scholar
  19. Samuelsson, B., 1972, Biosynthesis of prostaglandins, Fed. Proc. 31:1442–1450.PubMedGoogle Scholar
  20. Terragno, A., Rydzik, R., and Terragno, N., 1981, High performance liquid chromatography and UV detection for the separation and quantitation of prostaglandins, Prostaglandins 21:101–112.PubMedCrossRefGoogle Scholar
  21. van der Ouderaa, F.J., and Buytenhek, M., 1982, Purification of PGH synthase from sheep vesicular glands, Methods Enzymol. 86:60–68.PubMedCrossRefGoogle Scholar
  22. van der Ouderaa, F.J., Buytenhek, M., Nugteren, D.H., and van Dorp, D.A., 1977, Purification and characterisation of prostaglandin endoperoxide synthetase from sheep vesicular glands, Biochim. Biophys.Acta 487:315–331.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1985

Authors and Affiliations

  • Noreen J. Hickok
    • 1
  • Mary Alosio
    • 1
  • Richard S. Bookman
    • 1
  1. 1.Memorial Sloan-Kettering Cancer CenterNew YorkUSA

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