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. 1991 Apr 1;275(Pt 1):241–245. doi: 10.1042/bj2750241

Identification of a superoxide-generating NADPH oxidase system in human fibroblasts.

B Meier 1, A R Cross 1, J T Hancock 1, F J Kaup 1, O T Jones 1
PMCID: PMC1150038  PMID: 1850240

Abstract

Human fibroblasts have the capacity to release superoxide radicals upon stimulation of an electron transport system similar to the NADPH oxidase of leukocytes. Two components of the NADPH oxidase system, (1) a flavoprotein of 45 kDa which binds diphenylene iodonium (a compound described as a specific inhibitor of the leukocyte NADPH oxidase), and (2) a low-potential cytochrome b, are present in fibroblast membranes. Fibroblasts exhibit these compounds at lower concentrations than do polymorphonuclear leukocytes or B-lymphocytes. The superoxide-generating system is rather uniformly associated with the outer cell membrane, as shown by light and electron microscopy. Superoxide release upon stimulation with various agents was prevented by the addition of micromolar concentrations of diphenylene iodonium, making an NADPH oxidase a likely source.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adler S., Baker P. J., Johnson R. J., Ochi R. F., Pritzl P., Couser W. G. Complement membrane attack complex stimulates production of reactive oxygen metabolites by cultured rat mesangial cells. J Clin Invest. 1986 Mar;77(3):762–767. doi: 10.1172/JCI112372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Babior B. M., Curnutte J. T., McMurrich B. J. The particulate superoxide-forming system from human neutrophils. Properties of the system and further evidence supporting its participation in the respiratory burst. J Clin Invest. 1976 Oct;58(4):989–996. doi: 10.1172/JCI108553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Babior G. L., Rosin R. E., McMurrich B. J., Peters W. A., Babior B. M. Arrangement of the respiratory burst oxidase in the plasma membrane of the neutrophil. J Clin Invest. 1981 Jun;67(6):1724–1728. doi: 10.1172/JCI110210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beauchamp C., Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem. 1971 Nov;44(1):276–287. doi: 10.1016/0003-2697(71)90370-8. [DOI] [PubMed] [Google Scholar]
  5. Briggs R. T., Drath D. B., Karnovsky M. L., Karnovsky M. J. Localization of NADH oxidase on the surface of human polymorphonuclear leukocytes by a new cytochemical method. J Cell Biol. 1975 Dec;67(3):566–586. doi: 10.1083/jcb.67.3.566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Cross A. R. The inhibitory effects of some iodonium compounds on the superoxide generating system of neutrophils and their failure to inhibit diaphorase activity. Biochem Pharmacol. 1987 Feb 15;36(4):489–493. doi: 10.1016/0006-2952(87)90356-x. [DOI] [PubMed] [Google Scholar]
  9. Dorey C. K., Khouri G. G., Syniuta L. A., Curran S. A., Weiter J. J. Superoxide production by porcine retinal pigment epithelium in vitro. Invest Ophthalmol Vis Sci. 1989 Jun;30(6):1047–1054. [PubMed] [Google Scholar]
  10. Dutton P. L. Redox potentiometry: determination of midpoint potentials of oxidation-reduction components of biological electron-transfer systems. Methods Enzymol. 1978;54:411–435. doi: 10.1016/s0076-6879(78)54026-3. [DOI] [PubMed] [Google Scholar]
  11. Gatley S. J., Sherratt S. A. The effects of diphenyleneiodonium on mitochondrial reactions. Relation of binding of diphenylene[125I]iodonium to mitochondria to the extent of inhibition of oxygen uptake. Biochem J. 1976 Aug 15;158(2):307–315. doi: 10.1042/bj1580307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Görög P., Pearson J. D., Kakkar V. V. Generation of reactive oxygen metabolites by phagocytosing endothelial cells. Atherosclerosis. 1988 Jul;72(1):19–27. doi: 10.1016/0021-9150(88)90058-5. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hancock J. T., Maly F. E., Jones O. T. Properties of the superoxide-generating oxidase of B-lymphocyte cell lines. Determination of Michaelis parameters. Biochem J. 1989 Aug 15;262(1):373–375. doi: 10.1042/bj2620373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kunz W. S., Konstantinov A. A. Effect of b-c1-site inhibitors on the midpoint potentials of mitochondrial cytochromes b. FEBS Lett. 1983 May 8;155(2):237–240. doi: 10.1016/0014-5793(82)80611-x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Marcus A. J. Pathways of oxygen utilization by stimulated platelets and leukocytes. Semin Hematol. 1979 Jul;16(3):188–195. [PubMed] [Google Scholar]
  19. Marcus A. J. The role of lipids in platelet function: with particular reference to the arachidonic acid pathway. J Lipid Res. 1978 Sep;19(7):793–826. [PubMed] [Google Scholar]
  20. Matsubara T., Ziff M. Increased superoxide anion release from human endothelial cells in response to cytokines. J Immunol. 1986 Nov 15;137(10):3295–3298. [PubMed] [Google Scholar]
  21. Matsubara T., Ziff M. Superoxide anion release by human endothelial cells: synergism between a phorbol ester and a calcium ionophore. J Cell Physiol. 1986 May;127(2):207–210. doi: 10.1002/jcp.1041270203. [DOI] [PubMed] [Google Scholar]
  22. McCord J. M., Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969 Nov 25;244(22):6049–6055. [PubMed] [Google Scholar]
  23. Meier B., Radeke H. H., Selle S., Raspe H. H., Sies H., Resch K., Habermehl G. G. Human fibroblasts release reactive oxygen species in response to treatment with synovial fluids from patients suffering from arthritis. Free Radic Res Commun. 1990;8(3):149–160. doi: 10.3109/10715769009087988. [DOI] [PubMed] [Google Scholar]
  24. Meier B., Radeke H. H., Selle S., Younes M., Sies H., Resch K., Habermehl G. G. Human fibroblasts release reactive oxygen species in response to interleukin-1 or tumour necrosis factor-alpha. Biochem J. 1989 Oct 15;263(2):539–545. doi: 10.1042/bj2630539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murrell G. A., Francis M. J., Bromley L. Modulation of fibroblast proliferation by oxygen free radicals. Biochem J. 1990 Feb 1;265(3):659–665. doi: 10.1042/bj2650659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Murrell G. A., Francis M. J., Bromley L. Modulation of fibroblast proliferation by oxygen free radicals. Biochem J. 1990 Feb 1;265(3):659–665. doi: 10.1042/bj2650659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Radeke H. H., Meier B., Topley N., Flöge J., Habermehl G. G., Resch K. Interleukin 1-alpha and tumor necrosis factor-alpha induce oxygen radical production in mesangial cells. Kidney Int. 1990 Feb;37(2):767–775. doi: 10.1038/ki.1990.44. [DOI] [PubMed] [Google Scholar]
  28. Sawada M., Carlson J. C. Superoxide radical production in plasma membrane samples from regressing rat corpora lutea. Can J Physiol Pharmacol. 1989 May;67(5):465–471. doi: 10.1139/y89-074. [DOI] [PubMed] [Google Scholar]
  29. Sedor J. R., Carey S. W., Emancipator S. N. Immune complexes bind to cultured rat glomerular mesangial cells to stimulate superoxide release. Evidence for an Fc receptor. J Immunol. 1987 Jun 1;138(11):3751–3757. [PubMed] [Google Scholar]
  30. Shingu M., Yoshioka K., Nobunaga M., Motomatu T. C1q binding to human vascular smooth muscle cells mediates immune complex deposition and superoxide generation. Inflammation. 1989 Oct;13(5):561–569. doi: 10.1007/BF00916762. [DOI] [PubMed] [Google Scholar]
  31. Strittmatter C. F., Ball E. G. A Hemochromogen Component of Liver Microsomes. Proc Natl Acad Sci U S A. 1952 Jan;38(1):19–25. doi: 10.1073/pnas.38.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wach F., Hein R., Adelmann-Grill B. C., Krieg T. Inhibition of fibroblast chemotaxis by superoxide dismutase. Eur J Cell Biol. 1987 Aug;44(1):124–127. [PubMed] [Google Scholar]

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