Skip to main content
PMC home page
The Journal of Biophysical and Biochemical Cytology logoLink to The Journal of Biophysical and Biochemical Cytology
. 1957 May 25;3(3):397–412. doi: 10.1083/jcb.3.3.397

INTRACELLULAR LOCALIZATION OF ENZYMES IN SPLEEN

I. REDUCED DIPHOSPHOPYRIDINE NUCLEOTIDE CYTOCHROMEc REDUCTASE, CYTOCHROMEc OXIDASE, AND SUCCINIC DEHYDROGENASE IN THE RAT AND GUINEA PIG

Herbert J Eichel 1; With the Technical Assistance of Jeanne Hagan1
PMCID: PMC2224044  PMID: 13438924

Abstract

1. The intracellular distribution of nitrogen, DPNH cytochrome c reductase, succinic dehydrogenase, and cytochrome c oxidase has been studied in fractions derived by differential centrifugation from rat and guinea pig spleen homogenates. 2. In the spleens of each species, the nuclear fraction accounted for 40 to 50 per cent of the total nitrogen content of the homogenate, and the mitochondrial, microsome, and supernatant fractions contained about 8, 12, and 30 per cent of the total nitrogen, respectively. 3. Per mg. of nitrogen, DPNH cytochrome c reductase was concentrated in the mitochondria and microsomes of both rat and guinea pig spleens. Seventy per cent of the total DPNH cytochrome c reductase activity was recovered in these two fractions. The reductase activity associated with the nuclear fraction was lowered markedly by isolating nuclei from rat spleens with the sucrose-CaCl2 layering technique. The lowered activity was accompanied by the recovery of about 90 per cent of the homogenate DNA in the isolated nuclei, indicating that little, if any, of the reductase is present in spleen cell nuclei. 4. Per mg. of nitrogen, succinic dehydrogenase was concentrated about 10-fold in the mitochondria of rat spleen, and 65 per cent of the total activity was recovered in this fraction. 5. Cytochrome c oxidase was concentrated, per mg. of nitrogen, in the mitochondria of both rat and guinea pig spleens. The activity associated with the nuclear fraction was greatly diminished when this fraction was isolated from rat spleens by the sucrose-CaCl2 layering technique. Only 50 to 70 per cent of the total cytochrome c oxidase activity of the original homogenates was recovered among the four fractions from both rat and guinea pig spleens, while the specific activities of reconstructed homogenates were only 55 to 75 per cent of those of the original whole homogenates. This was in contrast to the results with DPNH cytochrome c reductase and succinic dehydrogenase where the recovery of total enzyme activity approached 100 per cent, and the specific activities of reconstructed homogenates equalled those of the original homogenates. The recovery of cytochrome c oxidase was greatly improved when only the nuclei were separated from rat spleen homogenates. 6. Data were presented comparing the concentrations (ratio of activity per mg. of nitrogen of the fraction to activity per mg. of nitrogen of the homogenate) of DPNH cytochrome c reductase in mitochondria and microsomes derived from different organs of different animals. 7. Data were presented comparing the activities per mg. of nitrogen of DPNH cytochrome c reductase in homogenates from several organs of various animals.

Full Text

The Full Text of this article is available as a PDF (952.1 KB).

Selected References

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

  1. BRODY T. M., WANG R. I. H., BAIN J. A. Intracellular distribution of diphosphopyridine nucleotide-cytochrome c reductase and cytochrome c oxidase in mammalian brain. J Biol Chem. 1952 Oct;198(2):821–826. [PubMed] [Google Scholar]
  2. DE DUVE C., PRESSMAN B. C., GIANETTO R., WATTIAUX R., APPELMANS F. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J. 1955 Aug;60(4):604–617. doi: 10.1042/bj0600604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. HOGEBOOM G. H. Cytochemical studies of mammalian tissues; the distribution of diphosphopyridine nucleotide-cytochrome c reductase in rat liver fractions. J Biol Chem. 1949 Feb;177(2):847–858. [PubMed] [Google Scholar]
  4. HOGEBOOM G. H., SCHNEIDER W. C. Intracellular distribution of enzymes. VIII. The distribution of diphosphopyridine nucleotidecytochrome C reductase in normal mouse liver and mouse hepatoma. J Natl Cancer Inst. 1950 Feb;10(4):983–987. [PubMed] [Google Scholar]
  5. HOGEBOOM G. H., SCHNEIDER W. C., STRIEBICH M. J. Cytochemical studies. V. On the isolation and biochemical properties of liver cell nuclei. J Biol Chem. 1952 May;196(1):111–120. [PubMed] [Google Scholar]
  6. JACKSON K. L., WALKER E. L., PACE N. Centrifugal preparation of rat liver mitochondria free of microsomes. Science. 1953 Jul 31;118(3057):136–137. doi: 10.1126/science.118.3057.136. [DOI] [PubMed] [Google Scholar]
  7. JACOB M., MANDEL L., MANDEL P. L'action du jeûne protéique prolongé sur les acides nucléiques de la rate et sa signification. Experientia. 1951 Jul 15;7(7):269–271. doi: 10.1007/BF02154549. [DOI] [PubMed] [Google Scholar]
  8. KUFF E. L. The distribution of fumarase activity in mouse liver homogenates. J Biol Chem. 1954 Mar;207(1):361–365. [PubMed] [Google Scholar]
  9. LAIRD A. K., NYGAARD O., RIS H., BARTON A. D. Separation of mitochondria into two morphologically and biochemically distinct types. Exp Cell Res. 1953 Sep;5(1):147–160. doi: 10.1016/0014-4827(53)90100-1. [DOI] [PubMed] [Google Scholar]
  10. LOMBARDO M. E., CERECEDO L. R., REDDY D. V. Nucleic acid changes during liver regeneration. J Biol Chem. 1953 May;202(1):97–106. [PubMed] [Google Scholar]
  11. MAXWELL E., ASHWELL G. Effect of x-irradiation on phosphorus metabolism in spleen mitochondria. Arch Biochem Biophys. 1953 Apr;43(2):389–398. doi: 10.1016/0003-9861(53)90134-9. [DOI] [PubMed] [Google Scholar]
  12. MUNTWYLER E., SEIFTER S., HARKNESS D. M. Some effects of restriction of dietary protein on the intracellular components of liver. J Biol Chem. 1950 May;184(1):181–190. [PubMed] [Google Scholar]
  13. NOVIKOFF A. B., PODBER E., RYAN J., NOE E. Biochemical heterogeneity of the cytoplasmic particles isolated from rat liver homogenate. J Histochem Cytochem. 1953 Jan;1(1):27–46. doi: 10.1177/1.1.27. [DOI] [PubMed] [Google Scholar]
  14. PETERMANN M. L., HAMILTON M. G. An ultracentrifugal analysis of the macromolecular particles of normal and leukemic mouse spleen. Cancer Res. 1952 May;12(5):373–378. [PubMed] [Google Scholar]
  15. POTTER V. R., RECKNAGEL R. O., HURLBERT R. B. Intracellular enzyme distribution; interpretations and significance. Fed Proc. 1951 Sep;10(3):646–653. [PubMed] [Google Scholar]
  16. RAMBACH W. A., MOOMAW D. R., ALT H. L., COOPER J. A. D. Nucleic acid metabolism of bone marrow and spleen. I. Normal values and effect of sodium pentobarbital. Proc Soc Exp Biol Med. 1952 Jan;79(1):59–62. doi: 10.3181/00379727-79-19272. [DOI] [PubMed] [Google Scholar]
  17. ROSE I. A., SCHWEIGERT B. S. Effect of vitamin B12 on nucleic acid metabolism of the rat. Proc Soc Exp Biol Med. 1952 Mar;79(3):541–544. doi: 10.3181/00379727-79-19439. [DOI] [PubMed] [Google Scholar]
  18. SCHNEIDER W. C., HOGEBOOM G. H. Intracellular distribution of succinoxidase and cytochrome oxidase activities in normal mouse liver and in mouse hepatoma. J Natl Cancer Inst. 1950 Feb;10(4):969–975. [PubMed] [Google Scholar]
  19. SHEPHERD J. A., KALNITSKY G. Intracellular distribution of fumarase, aconitase, and isocitric dehydrogenase in rabbit cerebral cortex. J Biol Chem. 1954 Apr;207(2):605–611. [PubMed] [Google Scholar]
  20. SHEPHERD J. A., LI Y. W., MASON E. E., ZIFFREN S. E. The distribution of aconitase and fumarase in homogenates of human liver. J Biol Chem. 1955 Mar;213(1):405–408. [PubMed] [Google Scholar]
  21. STRITTMATTER C. F., BALL E. G. The intracellular distribution of cytochrome components and of oxidative enzyme activity in rat liver. J Cell Physiol. 1954 Feb;43(1):57–78. doi: 10.1002/jcp.1030430105. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Biophysical and Biochemical Cytology are provided here courtesy of The Rockefeller University Press

RESOURCES