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. 1990 Sep 1;111(3):1217–1223. doi: 10.1083/jcb.111.3.1217

Purification of an inhibitor of erythroid progenitor cell cycling and antagonist to interleukin 3 from mouse marrow cell supernatants and its identification as cytosolic superoxide dismutase

PMCID: PMC2116302  PMID: 2391363

Abstract

We have isolated a protein from media conditioned by a murine marrow- derived cell line (PB6) and from mouse marrow supernatants that antagonizes interleukin 3-dependent proliferation of cells in culture and reversibly inhibits DNA synthesis of erythroid progenitor cells (BFU-E) in vitro. This protein, p16 (monomer Mr = 16 kD on SDS-PAGE), was purified to homogeneity and amino acid sequencing of a polypeptide fragment yielded a sequence identical to that of murine cytosolic Cu,Zn- containing superoxide dismutase (SOD). The identification of p16 as SOD was confirmed by the detection of SOD enzymatic activity in pure p16 fractions, and when a commercial human erythrocytic SOD preparation was tested it showed the same cell inhibitory activities as p16. These observations show that superoxide dismutase is able to affect the cycling and growth factor responses of hematopoietic cells, activities that have not previously been associated with this enzyme.

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Selected References

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  1. Axelrad A. A., Croizat H., Eskinazi D. A washable macromolecule from Fv2rr marrow negatively regulates DNA synthesis in erythropoietic progenitor cells BFU-E. Cell. 1981 Oct;26(2 Pt 2):233–244. doi: 10.1016/0092-8674(81)90306-8. [DOI] [PubMed] [Google Scholar]
  2. Axelrad A. A. Some hemopoietic negative regulators. Exp Hematol. 1990 Feb;18(2):143–150. [PubMed] [Google Scholar]
  3. 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]
  4. Beckman B. S., Balin A. K., Allen R. G. Superoxide dismutase induces differentiation of Friend erythroleukemia cells. J Cell Physiol. 1989 May;139(2):370–376. doi: 10.1002/jcp.1041390220. [DOI] [PubMed] [Google Scholar]
  5. Bewley G. C. cDNA and deduced amino acid sequence of murine Cu-Zn superoxide dismutase. Nucleic Acids Res. 1988 Mar 25;16(6):2728–2728. doi: 10.1093/nar/16.6.2728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark S. C., Kamen R. The human hematopoietic colony-stimulating factors. Science. 1987 Jun 5;236(4806):1229–1237. doi: 10.1126/science.3296190. [DOI] [PubMed] [Google Scholar]
  7. Del Rizzo D. F., Eskinazi D., Axelrad A. A. Negative regulation of DNA synthesis in early erythropoietic progenitor cells (BFU-E) by a protein purified from the medium of C57BL/6 mouse marrow cells. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4320–4324. doi: 10.1073/pnas.85.12.4320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Emerson S. G., Thomas S., Ferrara J. L., Greenstein J. L. Developmental regulation of erythropoiesis by hematopoietic growth factors: analysis on populations of BFU-E from bone marrow, peripheral blood, and fetal liver. Blood. 1989 Jul;74(1):49–55. [PubMed] [Google Scholar]
  9. Feldman L., Dainiak N. B-lymphocyte-derived erythroid burst-promoting activity is distinct from other known lymphokines. Blood. 1989 May 15;73(7):1814–1820. [PubMed] [Google Scholar]
  10. Fridovich I. Superoxide dismutases. Adv Enzymol Relat Areas Mol Biol. 1974;41(0):35–97. doi: 10.1002/9780470122860.ch2. [DOI] [PubMed] [Google Scholar]
  11. Kahr W. H., Lewis P. N., Pulleyblank D. E. H3 Cys-110 is in close proximity to the C-terminal regions of H2B and H4 in a nucleosome core with an altered internal arrangement of histones. Biochemistry. 1990 Jun 19;29(24):5821–5829. doi: 10.1021/bi00476a025. [DOI] [PubMed] [Google Scholar]
  12. Kensler T. W., Bush D. M., Kozumbo W. J. Inhibition of tumor promotion by a biomimetic superoxide dismutase. Science. 1983 Jul 1;221(4605):75–77. doi: 10.1126/science.6857269. [DOI] [PubMed] [Google Scholar]
  13. Marklund S. L. Extracellular superoxide dismutase and other superoxide dismutase isoenzymes in tissues from nine mammalian species. Biochem J. 1984 Sep 15;222(3):649–655. doi: 10.1042/bj2220649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Martin J. P., Jr, Dailey M., Sugarman E. Negative and positive assays of superoxide dismutase based on hematoxylin autoxidation. Arch Biochem Biophys. 1987 Jun;255(2):329–336. doi: 10.1016/0003-9861(87)90400-0. [DOI] [PubMed] [Google Scholar]
  15. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  16. 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]
  17. Meagher R. C., Salvado A. J., Wright D. G. An analysis of the multilineage production of human hematopoietic progenitors in long-term bone marrow culture: evidence that reactive oxygen intermediates derived from mature phagocytic cells have a role in limiting progenitor cell self-renewal. Blood. 1988 Jul;72(1):273–281. [PubMed] [Google Scholar]
  18. Oberley L. W., Buettner G. R. Role of superoxide dismutase in cancer: a review. Cancer Res. 1979 Apr;39(4):1141–1149. [PubMed] [Google Scholar]
  19. Oberley L. W., Ridnour L. A., Sierra-Rivera E., Oberley T. D., Guernsey D. L. Superoxide dismutase activities of differentiating clones from an immortal cell line. J Cell Physiol. 1989 Jan;138(1):50–60. doi: 10.1002/jcp.1041380109. [DOI] [PubMed] [Google Scholar]
  20. Pepinsky R. B. Localization of lipid-protein and protein-protein interactions within the murine retrovirus gag precursor by a novel peptide-mapping technique. J Biol Chem. 1983 Sep 25;258(18):11229–11235. [PubMed] [Google Scholar]
  21. Spitz D. R., Oberley L. W. An assay for superoxide dismutase activity in mammalian tissue homogenates. Anal Biochem. 1989 May 15;179(1):8–18. doi: 10.1016/0003-2697(89)90192-9. [DOI] [PubMed] [Google Scholar]
  22. Spivak J. L. The mechanism of action of erythropoietin. Int J Cell Cloning. 1986 May;4(3):139–166. doi: 10.1002/stem.5530040302. [DOI] [PubMed] [Google Scholar]
  23. Sporn M. B., Roberts A. B. Peptide growth factors are multifunctional. Nature. 1988 Mar 17;332(6161):217–219. doi: 10.1038/332217a0. [DOI] [PubMed] [Google Scholar]
  24. Suzuki S., Axelrad A. A. Fv-2 locus controls the proportion of erythropoietic progenitor cells (BFU-E) synthesizing DNA in normal mice. Cell. 1980 Jan;19(1):225–236. doi: 10.1016/0092-8674(80)90404-3. [DOI] [PubMed] [Google Scholar]
  25. Yu J., Shao L. E., Lemas V., Yu A. L., Vaughan J., Rivier J., Vale W. Importance of FSH-releasing protein and inhibin in erythrodifferentiation. Nature. 1987 Dec 24;330(6150):765–767. doi: 10.1038/330765a0. [DOI] [PubMed] [Google Scholar]

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