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Methods for the Determination of the Activity of Phospholipases and their Clinical Application

E. coli-based assay compared to a photometric micelle assay

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Esterases, Lipases, and Phospholipases

Part of the book series: NATO ASI Series ((NSSA,volume 266))

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Abstract

Many assays for the determination of phospholipase A2 activity [for review: 1] have been developed. The choice of a detection method depends partly on the purpose of a particular experiment. For example, some assays can be used on purified enzymes but are incompatible with crude systems, some methods provide a continuous assay and generate a time course while others do not, and some methods are amenable to automation while others are not. However, the most important consideration in the choice of the detection method is the sensitivity required for the particular enzyme which depends on the quantity of enzyme available and on its specific activity. This point is especially important for the assay of phospholipases A2 in human plasma, which are found in lower quantities and are, in general, less active than their counterparts from venom.

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References

  1. Reynolds, L.J., Washburn, W.N., Deems, R.A., and Dennis, E.A., Assay strategies and methods for phospholipases. Methods Enzym. 197: 1991; 3–23

    Article  CAS  Google Scholar 

  2. Crowl, R.M., Stoller, T.J., Conroy, R.R., and Stoner, C.R. Induction of phospholipase A2 gene expression in human hepatoma cells by mediators of the acute phase response. J. Biol. Chem. 266: 1991; 2647–2651

    PubMed  CAS  Google Scholar 

  3. Stefanski, E., Pruzanski, W., Sternby, B., and Vadas, P. Purification of a soluble phospholipase A2 from synovial fluid in rheumatoid arthritis. J. Biochem. Tokyo 100: 1986; 1297–1303

    PubMed  CAS  Google Scholar 

  4. Elsbach, P., Weiss, J., Wright, G.W.; Forst, S., van den Bergh, C.J., and Verheij, H.M. Regulation and role of phospholipases in host-bacteria interaction. Prog. Clin. Biol. Res. 349: 1990; 1–9

    PubMed  CAS  Google Scholar 

  5. Hoffmann, G.E., and Neumann, U. Modified photometric method for the determination of phospholipase A activities. Klin. Wochenschr. 67: 1989; 106–109

    Article  PubMed  CAS  Google Scholar 

  6. Elsbach, P., Weiss, J., Franson, R.C., Beckerdite-Quagliata, S., Scheider, A., and Harris, L. Separation and purification of a potent bactericidal/permeability-increasing protein and a closely associated phospholipase A2 from rabbit polymorphonuclear leukocytes. J. Biol. Chem. 254: 1979; 11000–11009

    PubMed  CAS  Google Scholar 

  7. Aufenanger, J., Zimmer, W., and Kattermann, R. Characteristics of a radiometric E.coli-based assay–Modified method for the determination of human serum phospholipase A, and its clinical application. Clin. Chem. 39: 1993; 605–613

    PubMed  CAS  Google Scholar 

  8. Dole, V.P. and Meinertz, H. Microdetermination of long-chain fatty acids in plasma and tissues. J. Biol. Chem. 235: 1969; 2595–2599

    Google Scholar 

  9. Durst, H.D., Milano, M., Kikta, E.J., Connelly, S.A., and Grushka, E. Phenylacyl esters of fatty acids via ether catalysts for enhanced ultraviolett detection in liquid chromatography. Anal. Chem. 41: 1975; 1797–1801

    Article  Google Scholar 

  10. Püttmann, M., Krug, H., v. Ochsenstein, E., and Kattermann, R. Determination of serum free fatty acids in the picomol range by an easy, fast and reliable high-performance liquid chromatography method. Clin. Chem. 39: 1993; 825–832

    PubMed  Google Scholar 

  11. Aufenanger, J. and Zimmer, W. 1993, Rapid isolation and purification of phospholipase A, from human serum submitted

    Google Scholar 

  12. Samman, M, Aufenanger, J. Püttmann, M., Zimmer, W., M. Quintel, and Katterman, R. Catalytically active pancreatic Phospholipase A2 as a prognostic marker in acute pancreatitis Gastroenterology: 1993 submitted

    Google Scholar 

  13. Märki, F., and Franson, R. Endogenous suppression of neutral-active and calcium-dependent phospholipase A2 in human polymorphonuclear leukocytes. Biochim. Biophys. Acta 879: 1986; 149–156

    Article  PubMed  Google Scholar 

  14. Waite, M. The Phospholipases. in: Handbook of Lipid Research, D J Hanahan (editor); Plenum Press, New York, Vol 5: 1987; 79–133

    Google Scholar 

  15. Parks, T.P., Lukas, S., and Hoffman, A.F. Purification and characterization of a phospholipase A2 from human osteoarthritic synovial fluid. in: Phospholipase A,,P.Y.K. Wong and E.A. Dennis (eds.), Plenum Press, New York, 1990; 55–81

    Google Scholar 

  16. Püttmann, M, Aufenanger, J., v. Ochsenstein, E., Dürholt, S., v. Ackern, K., Harenberg, J., and Hoffmann, G E Increased phospholipase A activities in sera of intensive-care patients show sn-2 specificity but no acyl-chain selectivity Clin. Chem. 39: 1993; 782–788

    Google Scholar 

  17. Schalkwijk, C.G., Märki, F., and Van den Bosch, H. Studies on the acyl-chain selectivty of cellular phospholipases A2. Biochim. Biophys. Acta 1044: 1990; 139–146

    Article  PubMed  CAS  Google Scholar 

  18. Elsbach, P., Weiss, J., and Kao, L. The role of intramembrane Ca’ in the hydrolysis of the phospholipids of Escherichia coli by Ca’-dependent phospholipases. J. Biol. Chem. 260: 1985; 1618–1622

    PubMed  CAS  Google Scholar 

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© 1994 Springer Science+Business Media New York

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Aufenanger, J., Samman, M., Quintel, M. (1994). Methods for the Determination of the Activity of Phospholipases and their Clinical Application. In: Mackness, M.I., Clerc, M. (eds) Esterases, Lipases, and Phospholipases. NATO ASI Series, vol 266. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-0993-0_31

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  • DOI: https://doi.org/10.1007/978-1-4899-0993-0_31

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4899-0995-4

  • Online ISBN: 978-1-4899-0993-0

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