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. 1989 Aug 15;262(1):373–375. doi: 10.1042/bj2620373

Properties of the superoxide-generating oxidase of B-lymphocyte cell lines. Determination of Michaelis parameters.

J T Hancock 1, F E Maly 1, O T Jones 1
PMCID: PMC1133273  PMID: 2554894

Abstract

The capacity of three B-lymphocyte cell lines to generate superoxide (O2.-) was examined. The Burkitt lymphoma lines P.3HR-1 and Jijoye gave no response to phorbol 12-myristate 13-acetate (PMA) at 100 ng/ml but produced up to 0.35 nmol of O2.-/min per mg of protein when stimulated with 5 micrograms of PMA/ml; the cell line RPMI 1788 produced Nitro Blue Tetrazolium-positive responses to low PMA concentrations and approx. 0.4 nmol of O2.-/min per mg of protein at 5 micrograms of PMA/ml. Each cell line contained approx. 10 pmol of low-potential cytochrome b (cytochrome b-245)/mg of protein. Homogenates of PMA-activated cells gave 10-20-fold greater rates of O2.- produced per mg of protein. The Km for NADPH varied between approx. 250 microM for P3.HR-1 and RPMI 1788 cell lines and 30.5 +/- 6.5 microM for the Jijoye cell line; the Km values for NADH were higher. Determination of intracellular NADPH concentration showed that this might limit the rate of O2.- production since in each cell line it was at or below the Km concentration.

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Selected References

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  1. Babior B. M. Oxygen-dependent microbial killing by phagocytes (first of two parts). N Engl J Med. 1978 Mar 23;298(12):659–668. doi: 10.1056/NEJM197803232981205. [DOI] [PubMed] [Google Scholar]
  2. Bravo R., Frank R., Blundell P. A., Macdonald-Bravo H. Cyclin/PCNA is the auxiliary protein of DNA polymerase-delta. Nature. 1987 Apr 2;326(6112):515–517. doi: 10.1038/326515a0. [DOI] [PubMed] [Google Scholar]
  3. Cross A. R., Jones O. T., Harper A. M., Segal A. W. Oxidation-reduction properties of the cytochrome b found in the plasma-membrane fraction of human neutrophils. A possible oxidase in the respiratory burst. Biochem J. 1981 Feb 15;194(2):599–606. doi: 10.1042/bj1940599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cross A. R., Jones O. T. The effect of the inhibitor diphenylene iodonium on the superoxide-generating system of neutrophils. Specific labelling of a component polypeptide of the oxidase. Biochem J. 1986 Jul 1;237(1):111–116. doi: 10.1042/bj2370111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cross A. R., Parkinson J. F., Jones O. T. Mechanism of the superoxide-producing oxidase of neutrophils. O2 is necessary for the fast reduction of cytochrome b-245 by NADPH. Biochem J. 1985 Mar 15;226(3):881–884. doi: 10.1042/bj2260881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cross A. R., Parkinson J. F., Jones O. T. The superoxide-generating oxidase of leucocytes. NADPH-dependent reduction of flavin and cytochrome b in solubilized preparations. Biochem J. 1984 Oct 15;223(2):337–344. doi: 10.1042/bj2230337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Halestrap A. P., Denton R. M. Specific inhibition of pyruvate transport in rat liver mitochondria and human erythrocytes by alpha-cyano-4-hydroxycinnamate. Biochem J. 1974 Feb;138(2):313–316. doi: 10.1042/bj1380313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hancock J. T., Jones O. T. The inhibition by diphenyleneiodonium and its analogues of superoxide generation by macrophages. Biochem J. 1987 Feb 15;242(1):103–107. doi: 10.1042/bj2420103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henderson L. M., Chappell J. B., Jones O. T. The superoxide-generating NADPH oxidase of human neutrophils is electrogenic and associated with an H+ channel. Biochem J. 1987 Sep 1;246(2):325–329. doi: 10.1042/bj2460325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Klebanoff S. J. Oxygen metabolism and the toxic properties of phagocytes. Ann Intern Med. 1980 Sep;93(3):480–489. doi: 10.7326/0003-4819-93-3-480. [DOI] [PubMed] [Google Scholar]
  11. Maly F. E., Cross A. R., Jones O. T., Wolf-Vorbeck G., Walker C., Dahinden C. A., De Weck A. L. The superoxide generating system of B cell lines. Structural homology with the phagocytic oxidase and triggering via surface Ig. J Immunol. 1988 Apr 1;140(7):2334–2339. [PubMed] [Google Scholar]
  12. Maly F. E., Nakamura M., Gauchat J. F., Urwyler A., Walker C., Dahinden C. A., Cross A. R., Jones O. T., de Weck A. L. Superoxide-dependent nitroblue tetrazolium reduction and expression of cytochrome b-245 components by human tonsillar B lymphocytes and B cell lines. J Immunol. 1989 Feb 15;142(4):1260–1267. [PubMed] [Google Scholar]
  13. Nakamura M., Murakami M., Koga T., Tanaka Y., Minakami S. Monoclonal antibody 7D5 raised to cytochrome b558 of human neutrophils: immunocytochemical detection of the antigen in peripheral phagocytes of normal subjects, patients with chronic granulomatous disease, and their carrier mothers. Blood. 1987 May;69(5):1404–1408. [PubMed] [Google Scholar]
  14. Pick E., Gadba R. Certain lymphoid cells contain the membrane-associated component of the phagocyte-specific NADPH oxidase. J Immunol. 1988 Mar 1;140(5):1611–1617. [PubMed] [Google Scholar]
  15. Royer-Pokora B., Kunkel L. M., Monaco A. P., Goff S. C., Newburger P. E., Baehner R. L., Cole F. S., Curnutte J. T., Orkin S. H. Cloning the gene for an inherited human disorder--chronic granulomatous disease--on the basis of its chromosomal location. Nature. 1986 Jul 3;322(6074):32–38. doi: 10.1038/322032a0. [DOI] [PubMed] [Google Scholar]
  16. Selvaraj R. J., Sbarra A. J. The role of the phagocyte in host-parasite interactions. VII. Di- and triphosphopyridine nucleotide kinetics during phagocytosis. Biochim Biophys Acta. 1967 Jul 25;141(2):243–249. doi: 10.1016/0304-4165(67)90097-9. [DOI] [PubMed] [Google Scholar]
  17. Volkman D. J., Buescher E. S., Gallin J. I., Fauci A. S. B cell lines as models for inherited phagocytic diseases: abnormal superoxide generation in chronic granulomatous disease and giant granules in Chediak-Higashi syndrome. J Immunol. 1984 Dec;133(6):3006–3009. [PubMed] [Google Scholar]

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