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. 1971 Oct;124(4):685–694. doi: 10.1042/bj1240685

The cell-free synthesis of cytochrome c by a microsomal fraction from rat liver

Néstor F González-Cadavid 1, Juan P Ortega 1, Magally González 1
PMCID: PMC1177245  PMID: 5131727

Abstract

Conditions were investigated for demonstrating the synthesis in vitro of the complete molecule of cytochrome c by isolated liver microsomal systems from partially hepatectomized rats. It was first found that in vivo the early labelled cytochrome c associated with the microsomal fraction required, by comparison with the mitochondrial pool, more drastic conditions of extraction and its binding was less affected by freezing and thawing of the subcellular particles. The procedure of extraction and purification of cytochrome c had to be modified accordingly, to assure the recovery of the recently synthesized molecule. Several subcellular fractions were isolated from regenerating liver with a homogenization medium containing either 5 or 10mm-Mg2+ and most of them were active in the synthesis of the cytochrome c apoprotein. The microsomal fraction, in the presence of either cell sap or pH5.0 fraction, was also able to incorporate [59Fe]haemin, δ-amino[3H]laevulic acid and 55Fe into the prosthetic group of cytochrome c. These experiments confirm firmly the conclusions of our previous results obtained in vivo showing that both the apoprotein and the haem moieties are made and linked together on cytoplasmic ribosomes and only then is the complete molecule transferred to the mitochondria.

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Selected References

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

  1. Aschenbrenner B., Druyan R., Albin R., Rabinowitz M. Haem a, cytochrome c and total protein turnover in mitochondria from rat heart and liver. Biochem J. 1970 Sep;119(2):157–160. doi: 10.1042/bj1190157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ashwell M., Work T. S. The biogenesis of mitochondria. Annu Rev Biochem. 1970;39:251–290. doi: 10.1146/annurev.bi.39.070170.001343. [DOI] [PubMed] [Google Scholar]
  3. BLOEMENDAL H., BONT W. S., BENEDETTI E. L. PREPARATION OF RAT-LIVER POLYSOMES WITHOUT THE UTILIZATION OF DETERGENT. Biochim Biophys Acta. 1964 May 18;87:177–180. doi: 10.1016/0926-6550(64)90064-7. [DOI] [PubMed] [Google Scholar]
  4. Beattie D. S., Basford R. E., Koritz S. B. Studies on the biosynthesis of mitochondrial protein components. Biochemistry. 1966 Mar;5(3):926–930. doi: 10.1021/bi00867a018. [DOI] [PubMed] [Google Scholar]
  5. Davidian N., Penniall R., Elliott W. B. Origin of mitochondrial enzymes. 3. Distribution and synthesis of cytochrome c in rat liver tissue. Arch Biochem Biophys. 1969 Sep;133(2):345–358. doi: 10.1016/0003-9861(69)90463-9. [DOI] [PubMed] [Google Scholar]
  6. González-Cadavid N. F., Bravo M., Campbell P. N. The significance of cytochrome c redistribution during the subcellular fractionation of rat liver. Biochem J. 1968 Apr;107(4):523–529. doi: 10.1042/bj1070523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. González-Cadavid N. F., Campbell P. N. Subcellular distribution of cytochrome c in rat liver. Methods for its extraction and purification. Biochem J. 1967 Nov;105(2):427–442. doi: 10.1042/bj1050427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. González-Cadavid N. F., Campbell P. N. The biosynthesis of cytochrome c. Sequence of incorporation in vivo of [14C]lysine into cytochrome c and total proteins of rat-liver subcellular fractions. Biochem J. 1967 Nov;105(2):443–450. doi: 10.1042/bj1050443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. González-Cadavid N. F., Wecksler M., Bravo M. Stimulating effect of haemin on the synthesis of cytochrome c by liver slices. FEBS Lett. 1970 Apr 16;7(3):248–250. doi: 10.1016/0014-5793(70)80172-7. [DOI] [PubMed] [Google Scholar]
  10. Haldar D., Freeman K., Work T. S. Biogenesis ommitochondria. Nature. 1966 Jul 2;211(5044):9–12. doi: 10.1038/211009a0. [DOI] [PubMed] [Google Scholar]
  11. Hawker J. S. Inhibition of sucrose phosphatase by sucrose. Biochem J. 1967 Feb;102(2):401–406. doi: 10.1042/bj1020401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kadenbach B. A quantitative study of the biosynthesis of cytochrome c. Eur J Biochem. 1969 Sep;10(2):312–317. doi: 10.1111/j.1432-1033.1969.tb00691.x. [DOI] [PubMed] [Google Scholar]
  13. Kadenbach B. Biosynthesis of cytochrome c. The sites of synthesis of apoprotein and holoenzyme. Eur J Biochem. 1970 Feb;12(2):392–398. doi: 10.1111/j.1432-1033.1970.tb00864.x. [DOI] [PubMed] [Google Scholar]
  14. Kadenbach B. Synthesis of mitochondrial proteins. The synthesis of cytochrome c in vitro. Biochim Biophys Acta. 1967 May 30;138(3):651–654. doi: 10.1016/0005-2787(67)90574-6. [DOI] [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. Morrison M., Horie S. Determination of heme a concentration in cytochrome preparations by hemochromogen method. Anal Biochem. 1965 Jul;12(1):77–82. doi: 10.1016/0003-2697(65)90144-2. [DOI] [PubMed] [Google Scholar]
  17. PATTERSON M. S., GREENE R. C. MEASUREMENT OF LOW ENERGY BETA-EMITTERS IN AQUEOUS SOLUTION BY LIQUID SCINTILLATION COUNTING OF EMULSIONS. Anal Chem. 1965 Jun;37:854–857. doi: 10.1021/ac60226a017. [DOI] [PubMed] [Google Scholar]
  18. PORTERFIELD J. S. A simple plaque-inhibition test for the study of arthropod-borne viruses. Bull World Health Organ. 1960;22:373–380. [PMC free article] [PubMed] [Google Scholar]
  19. Penniall R., Davidian N. Origin of mitochondrial enzymes I. Cytochrome c synthesis by endoplasmic reticulum. FEBS Lett. 1968 Jul;1(1):38–41. doi: 10.1016/0014-5793(68)80013-4. [DOI] [PubMed] [Google Scholar]
  20. ROODYN D. B., SUTTIE J. W., WORK T. S. Protein synthesis in mitochondria. 2. Rate of incorporation in vitro of radioactive amino acids into soluble proteins in the mitochondrial fraction, including catalase, malic dehydrogenase and cytochrome c. Biochem J. 1962 Apr;83:29–40. doi: 10.1042/bj0830029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Von Der Decken A., Campbell P. N. Studies on the synthesis of serum albumin by ribonucleoprotein particles isolated from rat liver. Biochem J. 1962 Sep;84(3):449–455. doi: 10.1042/bj0840449. [DOI] [PMC free article] [PubMed] [Google Scholar]

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