[en] In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared three methods of labeling it with 125Iodine:chloramine T, lactoperoxidase, and an original technique of 'self labeling' based on the ability of the enzyme to oxidize and bind 125I in the presence of H2O2. The chloramine T technique produced a degraded protein, as well shown by a high non-specific binding of tracer to antibody. The lactoperoxidase technique did not succeed in labeling MPO with an adequate specific activity. In contrast, the self-labeling method gave a stable tracer with a specific activity of 23 microCi/micrograms MPO (85 MBq), a satisfactory level of immunoreactivity, and a low-specific binding (less than or equal to 3%). After labeling, purification of tracer was achieved by gel filtration chromatography in phosphate buffer (0.05 M; pH7) to which 0.1% poly-L-lysine was added. The labeled molecule remained stable for 40 days and could be used for RIA with a polyclonal antibody raised in rabbits.
Disciplines :
Human health sciences: Multidisciplinary, general & others Anesthesia & intensive care
Author, co-author :
Deby, Ginette ; Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
Pincemail, Joël ; Centre Hospitalier Universitaire de Liège - CHU > Chirurgie cardio-vasculaire
Thirion, A.
Deby, C.
Lamy, Maurice ; Université de Liège - ULiège > Département des sciences cliniques > Anesthésie et réanimation
Franchimont, P.
Language :
English
Title :
Self-Labeling of Human Polymorphonuclear Leucocyte Myeloperoxidase with 125iodine
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Bibliography
Babior B.M. (1978) Oxygen-Dependent Microbial Killing by Phagocytes. New England Journal of Medicine 298:659.
Klebanoff S. (1968) Myeloperoxidase-halide-hydrogen peroxide antibacterial system. J Bacteriol 95:2131.
Janoff A., White R., Carp H., Harel S., Dearing R., Lee D. (1979) Lung injury induced by leukocytic proteases. Am J Pathol 97:111.
Bentwood B.J., Henson P.M. Immun. 1980, 124:855.
Agner K. Acta physiol. scand. 1941, 2:1.
Schultz J., Kaminker K. (1962) Myeloperoxidase of the leucocyte of normal human blood. I. Content and localization. Archives of Biochemistry and Biophysics 96:465.
Powe J.E., Short A., Sibbald W.J., Driedger A.A. (1982) Pulmonary accumulation of polymorphonuclear leukocytes in the adult respiratory distress syndrome. Critical Care Medicine 10:712.
Simon R.H., Ward P.A. Inflammation Basic Principles and Clinical Correlates , J. I., Gallin, I. M., Golstein, R., Snyderman, Raven Press, New York; 1988, 815.
Olofsson T., Olsson I., Venge P. Scand. J. Haemat. 1977, 18:73.
Venge P., Hällgren R., Stalenheim G., Olsson I. Scand. J. Haemat. 1979, 22:317.
Oberg G., Dahl R., Ellegaard J., Sundström C., Vaeth M., Venge P. Eur. J. Haemat. 1987, 38:148.
Olsen R.L., Steigen T.K., Holm T., Little C. (1986) Molecular forms of myeloperoxidase in human plasma. Biochem J 237:559.
Neumann S., Gunzer G., Lang H., Jochum M., Fritz H. (1986) Quantitation of myeloperoxidase from human granulocytes as an inflammation marker by enzyme-linked immunosorbent assay. Fresenius Z. analyt. Chem. 324:365.
Thorell J.I., Larsson I. (1971) Enzymatic iodination of polypeptides with 125I to high specific activity. Biochimica et Biophysica Acta (BBA) - Protein Structure 251:363.
Thorell J.I., Johansson B.G. (1974) Lactoperoxidase coupled to polyacrylamide for radio-iodination of proteins to high specific activity. Immunochemistry 11:203.
Greenwood F.C., Hunter W., Glover J. (1963) THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J 89:114.
Borgeat P., Samuelsson B. (1979) Arachidonic acid metabolism in polymorphonuclear leukocytes: Effects of ionophore A23187. Proceedings of the National Academy of Sciences 76:2148.
Bakkenist A.R.J., Wever R., Vulsma T., Plat H., Van Gelder B.F. (1978) Isolation procedure and some properties of myeloperoxidase from human leucocytes. Biochimica et Biophysica Acta (BBA) - Enzymology 524:45.
Agner K. (1958) Crystalline Myeloperoxidase. Acta Chemica Scandinavica 12:89.
Baught R.J., Travis J. (1976) Human leukocyte granule elastase: rapid isolation and characterization. Biochemistry 15:836.
Brewer J.M., Ashworth R.B. (1969) Disc electrophoresis. Journal of Chemical Education 46:41.
Segrest J.P., Jackson R.L. (1972) [5] Molecular weight determination of glycoproteins by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Meth. Enzym. 28:54.
Wood W.G., Stella G., Müller O.A., Seriba P.C. (1979) A rapid and specific method for separation of bound and free antigen in radioimmunoassay systems. J Clin Chem Clin Biochem 17:111.
Pincemail J., Deby-Dupont G., Deby C., Thirion A., Torpier G., Faymonville M.E., Damas P., Tomassini M., Lamy M., Franchimont P. (1991) Fast double antibody radioimmunoassay of human granolocyte myeloperoxidase and its application to plasma. Journal of Immunological Methods 137:181.
Matheson N.R. (1982) The effect of antiarthritic drugs and related compounds on the human neutrophil myeloperoxidase system. Biochemical and Biophysical Research Communications 108:259.
Wong P.S., Travis J. (1980) Isolation and properties of oxidized alpha-1-proteinase inhibitor from human rheumatoid synovial fluid. Biochemical and Biophysical Research Communications 96:1449.
Matheson N.R., Wong P.S., Travis J. (1981) Isolation and properties of human neutrophil myeloperoxidase. Biochemistry 20:325.
Andrews P.C., Krinsky N.I. (1981) The reductive cleavage of myeloperoxidase in half, producing enzymically active hemi-myeloperoxidase. J Biol Chem 256:4211.
Harrison J.E., Pablan S., Schultz J. (1977) The subunit structure of crystalline canine myeloperoxidase. Biochimica et Biophysica Acta (BBA) - Protein Structure 493:247.
Andersen M.R., Atkin C.L., Eyre H.J. (1982) Intact form of myeloperoxidase from normal human neutrophils. Archives of Biochemistry and Biophysics 214:273.
Nauseef W., Root R.K., Malech H.L. (1983) Biochemical and Immunologic Analysis of Hereditary Myeloperoxidase Deficiency. Journal of Clinical Investigation 71:1297.
Olsen R.L., Little C. (1984) Studies on the subunits of human myeloperoxidase. Biochem J 222:701.
Olsson I., Olofsson T., Odeberg H. Scand. J. Haemat. 1972, 9:483.
Svensson B.E., Domeij K., Lindvall S., Rydell G. (1987) Peroxidase and peroxidase-oxidase activities of isolated human myeloperoxidases. Biochem J 242:673.
Olsson I. Eur. J. clin. Invest. 1978, 17:322.
Arnljots K., Olsson I. (1987) Myeloperoxidase precursors incorporate heme. J Biol Chem 262:10430.
Zgliezynski M., Stelmzynska T., Domanski J., Ostrowski W. (1971) Chloramines as intermediates of oxidation reaction of amino acids by myeloperoxidase. Biochimica et Biophysica Acta (BBA) - Enzymology 235:419.
Hager L.P., Morris D.R., Brown F.S., Eberwein H. (1966) Chloroperoxidase. II. Utilization of halogen anions. J Biol Chem 241:1769.
Klebanoff S.J. (1967) IODINATION OF BACTERIA: A BACTERICIDAL MECHANISM. Journal of Experimental Medicine 126:1063.
Klebanoff S.J., Clark R.A. J. Labo. clin. Med. 1977, 89:675.
Odijama T., Yamazaki I. Biochem. biophys. Act. 1970, 206:71.
Pincemail J., Faymonville M.E., Lamy M. Update in Intensive Care and Emergency Medicine, vol. 8 , J. L., Vincent, Springer-Verlag, Berlin; 1989, 24-32.
.
Pincemail J., Camus G., Roesgen A., Dreezen E., Bertrand Y., Lismonde M., Deby-Dupont G., Deby C. (1990) Exercise induces pentane production and neutrophil activation in humans. Effect of propranolol. European Journal of Applied Physiology and Occupational Physiology 61:319.
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