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. 1979 Oct 1;83(1):231–239. doi: 10.1083/jcb.83.1.231

Membrane-bound redox proteins of the murine Friend virus-induced erythroleukemia cell

PMCID: PMC2110442  PMID: 292645

Abstract

We have obtained and studied a 105,000-g pellet from T-3-Cl-2 cells, a cloned line of Friend virus-induced erythroleukemia cells. By difference spectrophotometry, the pellet was shown to contain cytochrome b5 and cytochrome P-450, hemeproteins that have been shown to participate in electron-transport reactions of endoplasmic reticulum and other membranous fractions of various tissues. The pellet also possesses NADH-cytochrome c reductase activity which is inhibited by anti-cytochrome b5 gamma-globulin, indicating the presence of cytochrome b5 reductase. This is the first demonstration of membrane- bound forms of these redox proteins in erythroid cells. Dimethyl sulfoxide-treated T-3-Cl-2 cells were also shown to possess membrane- bound cytochrome b5 and NADH-cytochrome c reductase activity. We failed to detect soluble cytochrome b5 in the 105,000-g supernatant fraction from homogenates of untreated or dimethyl sulfoxide-treated T-3-Cl-2 cells. In contrast, erythrocytes obtained from mouse blood were shown to possess soluble cytochrome b5 but no membrane-bound form of this protein. These findings are supportive of our hypothesis that soluble cytochrome b5 of erythrocytes is derived from endoplasmic reticulum or some other membrane structure of immature erythroid cells during cell maturation.

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Selected References

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  1. ACKERMAN G. A., GRASSO J. A., KNOUFF R. A. Erythropoiesis in the mammalian embryonic liver as revealed by electron microscopy. Lab Invest. 1961 Jul-Aug;10:787–796. [PubMed] [Google Scholar]
  2. Berezney R., Macaulay L. K., Crane F. L. The purification and biochemical characterization of bovine liver nuclear membranes. J Biol Chem. 1972 Sep 10;247(17):5549–5561. [PubMed] [Google Scholar]
  3. Boyer S. H., Wuu K. D., Noyes A. N., Young R., Scher W., Friend C., Preisler H. D., Bank A. Hemoglobin biosynthesis in murine virus-induced leukemic cells in vitro: structure and amounts of globin chains produced. Blood. 1972 Dec;40(6):823–835. [PubMed] [Google Scholar]
  4. Brownie A. C., Simpson E. R., Skelton F. R., Elliott W. B., Estabrook R. W. Interaction of androgens with the adrenal mitochondrial cytochrome system. The influence of androgen treatment on the levels of cytochrome in rat adrenal cortical mitochondria and on substrate-induced spectral changes. Arch Biochem Biophys. 1970 Nov;141(1):18–25. doi: 10.1016/0003-9861(70)90101-3. [DOI] [PubMed] [Google Scholar]
  5. Capalna S. The erythrocyte cytochrome b5. Physiologie. 1977 Apr-Jun;14(2):85–87. [PubMed] [Google Scholar]
  6. Douglas R. H., Hultquist D. E. Evidence that two forms of bovine erythrocyte cytochrome b5 are identical to segments of microsomal cytochrome b5. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3118–3122. doi: 10.1073/pnas.75.7.3118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. FRASCA J. M., PARKS V. R. A ROUTINE TECHNIQUE FOR DOUBLE-STAINING ULTRATHIN SECTIONS USING URANYL AND LEAD SALTS. J Cell Biol. 1965 Apr;25:157–161. doi: 10.1083/jcb.25.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fowler S., Remacle J., Trouet A., Beaufay H., Berthet J., Wibo M., Hauser P. Analytical study of microsomes and isolated subcellular membranes from rat liver. V. Immunological localization of cytochrome b5 by electron microscopy: methodology and application to various subcellular fractions. J Cell Biol. 1976 Nov;71(2):535–550. doi: 10.1083/jcb.71.2.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Franke W. W., Deumling B., Baerbelermen, Jarasch E. D., Kleinig H. Nuclear membranes from mammalian liver. I. Isolation procedure and general characterization. J Cell Biol. 1970 Aug;46(2):379–395. doi: 10.1083/jcb.46.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Friend C., Patuleia M. C., De Harven E. Erythrocytic maturation in vitro of murine (Friend) virus-induced leukemic cells. Natl Cancer Inst Monogr. 1966 Sep;22:505–522. [PubMed] [Google Scholar]
  11. Hultquist D. E., Dean R. T., Douglas R. H. Homogeneous cytochrome b5 from human erythrocytes. Biochem Biophys Res Commun. 1974 Sep 9;60(1):28–34. doi: 10.1016/0006-291x(74)90167-3. [DOI] [PubMed] [Google Scholar]
  12. Ikawa Y., Furusawa M., Sugano H. Erythrocyte membrane-specific antigens in Friend virus-induced leukemia cells. Bibl Haematol. 1973;39:955–967. doi: 10.1159/000427928. [DOI] [PubMed] [Google Scholar]
  13. Kasper C. B. Biochemical distinctions between the nuclear and microsomal membranes from rat hepatocytes. J Biol Chem. 1971 Feb 10;246(3):577–581. [PubMed] [Google Scholar]
  14. Kuma F., Inomata H. Studies on methemoglobin reductase. II. The purification and molecular properties of reduced nicotinamide adenine dinucleotide-dependent methemoglobin reductase. J Biol Chem. 1972 Jan 25;247(2):556–560. [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Lau A. F., Ruddon R. W. Proteins of transcriptionally active and inactive chromatin from Friend erythroleukemia cells. Exp Cell Res. 1977 Jun;107(1):35–46. doi: 10.1016/0014-4827(77)90383-4. [DOI] [PubMed] [Google Scholar]
  17. Levine E. M. Mycoplasma contamination of animal cell cultures: a simple, rapid detection method. Exp Cell Res. 1972 Sep;74(1):99–109. doi: 10.1016/0014-4827(72)90484-3. [DOI] [PubMed] [Google Scholar]
  18. MASSEY V. The microestimation of succinate and the extinction coefficient of cytochrome c. Biochim Biophys Acta. 1959 Jul;34:255–256. doi: 10.1016/0006-3002(59)90259-8. [DOI] [PubMed] [Google Scholar]
  19. OMURA T., SATO R. THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE. J Biol Chem. 1964 Jul;239:2370–2378. [PubMed] [Google Scholar]
  20. Orkin S. H., Harosi F. I., Leder P. Differentiation in erythroleukemic cells and their somatic hybrids. Proc Natl Acad Sci U S A. 1975 Jan;72(1):98–102. doi: 10.1073/pnas.72.1.98. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ostertag W., Melderis H., Steinheider G., Kluge N., Dube S. Synthesis of mouse haemoglobin and globin mRNA in leukaemic cell cultures. Nat New Biol. 1972 Oct 25;239(95):231–234. doi: 10.1038/newbio239231a0. [DOI] [PubMed] [Google Scholar]
  22. Passon P. G., Hultquist D. E. Soluble cytochrome b 5 reductase from human erythrocytes. Biochim Biophys Acta. 1972 Jul 12;275(1):62–73. doi: 10.1016/0005-2728(72)90024-2. [DOI] [PubMed] [Google Scholar]
  23. Passon P. G., Reed D. W., Hultquist D. E. Soluble cytochrome b 5 from human erythrocytes. Biochim Biophys Acta. 1972 Jul 12;275(1):51–61. doi: 10.1016/0005-2728(72)90023-0. [DOI] [PubMed] [Google Scholar]
  24. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Remacle J. Binding of cytochrome b5 to membranes of isolated subcellular organelles from rat liver. J Cell Biol. 1978 Nov;79(2 Pt 1):291–313. doi: 10.1083/jcb.79.2.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ross J., Ikawa Y., Leder P. Globin messenger-RNA induction during erythroid differentiation of cultured leukemia cells. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3620–3623. doi: 10.1073/pnas.69.12.3620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. STRITTMATTER P., VELICK S. F. A microsomal cytochrome reductase specific for diphosphopyridine nucleotide. J Biol Chem. 1956 Jul;221(1):277–286. [PubMed] [Google Scholar]
  28. Sato T., Friend C., De Harven E. Ultrastructural changes in Friend erythroleukemia cells treated with dimethyl sulfoxide. Cancer Res. 1971 Oct;31(10):1402–1417. [PubMed] [Google Scholar]
  29. Sottocasa G. L., Kuylenstierna B., Ernster L., Bergstrand A. An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study. J Cell Biol. 1967 Feb;32(2):415–438. doi: 10.1083/jcb.32.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sugita Y., Nomura S., Yoneyama Y. Purification of reduced pyridine nucleotide dehydrogenase from human erythrocytes and methemoglobin reduction by the enzyme. J Biol Chem. 1971 Oct 10;246(19):6072–6078. [PubMed] [Google Scholar]
  31. Weibel E. R., Stäubli W., Gnägi H. R., Hess F. A. Correlated morphometric and biochemical studies on the liver cell. I. Morphometric model, stereologic methods, and normal morphometric data for rat liver. J Cell Biol. 1969 Jul;42(1):68–91. doi: 10.1083/jcb.42.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zbarsky I. B., Perevoshchikova K. A., Delektorskaya L. N., Delektorsky V. V. Isolation and biochemical characteristics of the nuclear envelope. Nature. 1969 Jan 18;221(5177):257–259. doi: 10.1038/221257a0. [DOI] [PubMed] [Google Scholar]

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