, Volume 10, Issue 2, pp 183–193 | Cite as

Influence of plasma proteins on activity of proinflammatory enzyme phospholipase A2

  • P. Vadas
  • E. Stefanski
  • W. Pruzanski
Original Articles


The influence of plasma proteins on quantitation of the proinflammatory enzyme phospholipase A2 (PLA2) from rheumatoid synovial fluid was studied using two different assays. Human and bovine serum albumins increased the rate of PLA, hydrolysis of membrane-associated phospholipid substrates. In contrast, albumin profoundly inhibited the PLA2 hydrolysis of synthetic phospholipids in micellar dispersion. Other plasma proteins (α,β andγ-globulins) had minimal effect on PLA2 activity in either assay system. Since the presence of albumin may compromise estimation of PLA2 and of phospholipase-inhibitory proteins, the appropriate selection of assay conditions is obligatory for the accurate quantitation of their respective activities.


Public Health Internal Medicine Albumin Bovine Serum Bovine Serum Albumin 
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  1. 1.
    Vadas, P., andJ. Hay. 1980. The release of phospholipase A2 from aggregated platelets and stimulated macrophages of sheep.Life Sci. 26:1721–1729.Google Scholar
  2. 2.
    Vadas, P., S. Wasi, M. Movat, andJ. Hay. 1981. Extracellular phospholipase A2 mediates inflammatory hyperemia.Nature 293:583–585.Google Scholar
  3. 3.
    Wightman, P., M. Dahlgren, P. Davis, andR. Bonney. 1981. The selective release of phospholipase A2 by resident mouse peritoneal macrophages.Biochem. J. 200:441–444.Google Scholar
  4. 4.
    Traynor, M., andK. Authi. 1981. Phospholipase A2 activity of lysosomal origin secreted by polymorphonuclear leukocytes during phagocytosis or on treatment with calcium.Biochim. Biophys. Acta. 665:571–577.Google Scholar
  5. 5.
    Lanni, C., andE. Becker. 1983. Release of phospholipase A2 activity from rabbit peritoneal neutrophils by f-met-leu-phe.Am. J. Pathol. 113:90–94.Google Scholar
  6. 6.
    Vadas, P., andW. Pruzanski. 1984. Role of extracellular phospholipase A2 in inflammation.Adv. Inflam. Res. 7:51–59.Google Scholar
  7. 7.
    Vadas, P., andJ. Hay. 1982. The appearance and significance of phospholipase A2 in lymph draining tuberculin reactions.Am. J. Pathol. 107:285–291.Google Scholar
  8. 8.
    Franson, R., R. Doblow, J. Weiss, P. Elsbach, andW. Weglicki. 1978. Isolation and characterization of a phospholipase A2 from an inflammatory exudate.J. Lipid Res. 19:18–23.Google Scholar
  9. 9.
    Nevalainen, T. 1980. The role of phospholipase A in acute pancreatitis.Scand. J. Gastroenterol. 15:641–650.Google Scholar
  10. 10.
    Pruzanski, W., P. Vadas, E. Stefanski, andM. Urowitz. 1985. Phospholipase A2 activity in sera and synovial fluids in rheumatoid arthritis and osteoarthritis. Its possible role as a proinflammatory enzyme.J. Rheumatol. 12:211–216.Google Scholar
  11. 11.
    Vadas, P., E. Stefanski, andW. Pruzanski. 1985. Characterization of extracellular phospholipase A2 in rheumatoid synovial fluid.Life Sci. 36:579–588.Google Scholar
  12. 12.
    Shaw, J., M. Roberts, R. Ulevitch, P. Henson, andE. Dennis. 1978. Phospholipase A2 contamination of cobra venom factor preparations.Am. J. Pathol. 91:517–530.Google Scholar
  13. 13.
    Pruzanski, W., P.Vadas, and V.Fornasier. 1986. Inflammatory effect of intradermal administration of soluble phospholipase A2 in rabbits.J. Invest. Dermatol. (in press).Google Scholar
  14. 14.
    Blackwell, G., R. Carnuccio, M. DiRosa, R. Flower, C. Langham, L. Parente, P. Persico, N. Russel-Smith, andD. Stone. 1982. Glucoconicoids induce the formation and release of antiinflammatory and antiphospholipase proteins into the peritoneal cavity of the rat.Br. J. Pharmacol. 76:185–194.Google Scholar
  15. 15.
    Parente, L., M. DiRosa, R. Flower, P. Ghiara, R. Meli, P. Persico, J. Salmon, andJ. Wood. 1984. Relationship between the antiphospholipase and antiinflammatory effects of glucocorticoid-induced proteins.Eur. J. Pharmacol. 99:233–239.Google Scholar
  16. 16.
    Hirata, F., E. Schiffman, K. Venkatasubramanian, D. Salomon, andJ. Axelrod. 1980. A phospholipase A2 inhibitory protein in rabbit neutrophils induced by glucocorticoids.Proc. Natl. Acad. Sci. U.S.A. 77:2533–2536.Google Scholar
  17. 17.
    Cloix, J., O. Colard, B. Rothhut, andF. Russo-Marie. 1983. Characterization and partial purification of renocortins; two polypeptides formed in renal cells causing the antiphospholipase-like action of glucocorticoids.Br. J. Pharmacol. 79:313–321.Google Scholar
  18. 18.
    Weglicki, W., R. Ruth, K. Owens, H. Griffith, andM. Waite. 1974. Changes in lipid composition of Triton-filled lysosomes during lysis.Biochim. Biophys. Acta 337:145–152.Google Scholar
  19. 19.
    Lin, H-K., andE. J. Simon. 1978. Phospholipase A2 inhibition of opiate receptor binding can be reversed by albumin.Nature 271:383–384.Google Scholar
  20. 20.
    Hirata, F., Y. Notsu, M. Iwata, L. Parente, M. DiRosa, andR. Flower. 1982. Identification of several species of phospholipase inhibitory protein(s) by radioimmunoassay for lipomodulin.Biochem. Biophys. Res. Commun. 109:223–230.Google Scholar
  21. 21.
    Heath, M., andW. Jacobson. 1984. The effect of components of rabbit pulmonary surfactant on the activity of phospholipases.J. Physiol. 346:439–448.Google Scholar
  22. 22.
    Shakir, K. M. 1981. Phospholipase A2 activity of postheparin plasma: A rapid and sensitive assay and partial characterization.Anal. Biochem. 114:64–70.Google Scholar
  23. 23.
    Nishijima, J., M. Okamoto, M. Ogawa, G. Kosaki, andJ. Yamano. 1983. Purification and characterization of human pancreatic phospholipase A2 and development of a radioimmunoassay.J. Biochem. 94:137–147.Google Scholar
  24. 24.
    Rubin, A. L., G. D. Lubash, R. Aronson, andP. Davison. 1963. Separation of polypeptides bound by albumin in human plasma.Nature 197:1009–1010.Google Scholar
  25. 25.
    Schmid, K., andM. B. MacNair. 1956. Characterization of the proteins of human synovial fluid in certain disease states.J. Clin. Invest. 35:814–824.Google Scholar
  26. 26.
    Van Den Bosch, J., andA. J. Aarsman.1979. A review on methods of phospholipase A determination.Agents Actions 9:382–389.Google Scholar
  27. 27.
    Dennis, E. A. 1973. Kinetic dependence of phospholipase A2 activity on the detergent Triton X-100.J. Lipid Res. 14:152–159.Google Scholar
  28. 28.
    Franson, R., andM. Waite. 1978. Relationship between calcium requirement, substrate charge, and rabbit polymorphonuclear phospholipase A2 activity.Biochemistry 17:4029–4033.Google Scholar
  29. 29.
    Van Den Bosch, H. 1980. Intracellular phospholipases A.Biochim. Biophys. Acta 604:191–246.Google Scholar
  30. 30.
    Spector, A. A., J. Fletcher, andJ. D. Ashbrook. 1971. Analysis of long-chain free fatty acid binding to bovine serum albumin by determination of stepwise equilibrium constants.Biochemistry 10:3229–3232.Google Scholar
  31. 31.
    Sweet, C., andJ. E. Zull. 1970. The binding of serum albumin to phospholipid liposomes.Biochim. Biophys. Acta 219:253–262.Google Scholar
  32. 32.
    Smith, A. D., S. Gul, andR. H. S. Thompson, 1972. The effect of fatty acids and of albumin on the action of a purified phospholipase A2 from cobra venom on synthetic lecithins.Biochim. Biophys. Acta 289:147–157.Google Scholar
  33. 33.
    Jonas, A. 1975. Phosphatidyl choline interaction with bovine serum albumin: Effect of the physical state of the lipid on protein-lipid complex formation.Biochem. Biophys. Res. Commun. 64:1003–1008.Google Scholar
  34. 34.
    Nevalainen, T., andO. Evilampi. 1984. Presence of a phospholipase A2 inhibitor in porcine serum.J. Biochem. 96:1303–1305.Google Scholar
  35. 35.
    Etienne, J., A. Gruber, andJ. Polonovski. 1984. Inhibitory factor of phospholipase A2 in normal human serum.Biochem. Biophys. Res. Commun. 122:1117–1124.Google Scholar
  36. 36.
    Miwa, M., I. Kubota, T. Ichihashi, H. Motojima, andM. Matsumoto. 1984. Studies on phospholipase A inhibitor in blood plasma. I. Purification and characterization of phospholipase A inhibitor in bovine plasma.J. Biochem. 96:761–773.Google Scholar
  37. 37.
    Hirata, F. 1981. The regulation of lipomodulin, a phospbolipase-inhibitory protein, in rabbit neutrophils by phosphorylation.J. Biol. Chem. 256:7730–7733.Google Scholar
  38. 38.
    Iwata, M., T. Huff, andK. Ishizaka. 1984. Modulation of the biologic activities of IgE-binding factor.J. Immunol. 132:1286–1293.Google Scholar
  39. 39.
    Hirata, F. 1983. Lipomodulin: A possible mediator of the action of glucocorticoids.In Advances in Prostaglandin, Thromboxanes and Leukotriene Research, Vol. 2. B. Samuelsson. R. Paoletti, and P. Ramwell, editors. Raven Press, New York. 73–78.Google Scholar
  40. 40.
    Flower, R. J. 1984. Macrocortin and the antiphospholipase proteins.Adv. Inflam. Res. 8:1–34.Google Scholar
  41. 41.
    Franson, R., P. Patriarca, andP. Elsbach. 1974. Phospholipid metabolism by phagocytic cells. Phospholipases A2 associated with polymorphonuclear leukocyte granules.J. Lipid Res. 15:380–388.Google Scholar

Copyright information

© Plenum Publishing Corporation 1986

Authors and Affiliations

  • P. Vadas
    • 1
  • E. Stefanski
    • 1
  • W. Pruzanski
    • 1
  1. 1.Immunology Diagnostic and Research Centre Rheumatic Diseases Unit, Department of Medicine The Wellesley HospitalUniversity of TorontoTorontoCanada

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