Skip to main content
Log in

Studies of adenosine incorporation in Langendorff rat heart and rat heart mitochondria

  • Original Article
  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The acid-insoluble product isolated from well-oxygenated Langendorff rat heart after perfusion with [14C]adenosine was purified by phenol extraction and subjected to specific phosphorolysis by pure polynucleotide phosphorylase. TLC analysis of the reaction mixture showed that ADP was the only radioactive product, proving that the original substance was a polyribonucleotide. Studies of the time course of labelling and of the distribution of the acid-insoluble product between the mitochondrial and nuclear fractions showed that both are labelled even after 1 min at 25 °C, but at short times and low temperature more radioactivity is found in the mitochondria. The kinetics of adenosine incorporation resemble those expected for the labelling of hnRNA and mRNA. Isolated, respiring mitochondria incorporate adenosine and adenine nucleotides into acid insoluble form by a process dependent on oxidative phosphorylation and the adenine nucleotide translocase that is specific for adenine derivatives. The results are discussed in terms of the hypothesis that the polyribonucleotide might be a storage form of adenine nucleotides: it is concluded that the bulk of the labelled product is unlikely to play a major role in energy metabolism.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Swain JL, Sabina RL, McHale PA, Greenfield Jr JC, Holmes EW: Derangements in myocardial purine and pyrimidine nucleotide metabolism in patients with coronary artery disease and left ventricular hypertrophy. Amer J Physiol 242:H818-H826, 1982

    Google Scholar 

  2. Jennings RB, Hawkins HK, Lowe JE, Hill ML, Klotman S, Reimer KA: Relation between high energy phosphate and lethal injury in myocardial ischemia in the dog. Amer J Pathol 92:187–214, 1978

    Google Scholar 

  3. Swain JL, Holmes EW: Nucleotide metabolism in cardiac muscle. In: HA Fozzard (ed) The Heart and Cardiovascular System. Raven Press, New York, 1986, pp 911–929

    Google Scholar 

  4. Young HH, Shimizu T, Nishioka K, Nakenishi T, Jarmakani JM: Effect of hypoxia and reoxygenation on mitochondrial function in neonatal myocardium. Amer J Physiol 245:H998-H1006, 1983

    Google Scholar 

  5. Watanabe F, Kamiiki W, Nishimura T, Hashimoto T, Tagawa K: Decrease in mitochondrial levels of adenine nucleotides and concomitant mitochondrial disfunction in ischemic rat liver. J. Biochem 94:493–499, 1983

    Google Scholar 

  6. Takeo S, Sakanashi M: Possible mechanisms for reoxygenation-induced recovery of myocardial high-energy phosphates after hypoxia. J. Mol Cell Cardiol 15:577–594, 1983

    Google Scholar 

  7. Hutchinson WL, Bates DJ, Perrett D, Mowbray J: In perfused rat hearts ischaemia promotes the reversible conversion of appreciable quantities of soluble adenine nucleotides to a stable trichloroacetic acid-precipitable form. Biosci Rep 1:857–863, 1981

    Google Scholar 

  8. Mowbray J, Hutchinson WL, Tibbs GR, Morris PG: The discovery of a rapidly metabolized polymeric tetraphosphate derivative of adenosine in perfused rat heart. Biochem J 223:627–632, 1984

    Google Scholar 

  9. Hutchinson WL, Morris PG, Mowbray J: The molecular structure of a rapidly formed oligomeric adenosine tetraphosphate derivative from rat heart. Biochem J 234:623–627, 1986

    Google Scholar 

  10. Zimmer HG and Ibel H: Ribose accelerates the repletion of the ATP pool during recovery from reversible ischemia of the rat myocardium. J Mol Cell Cardiol 16:863–866, 1984

    Google Scholar 

  11. Fitt PS, Korecky B, Sharma N: A possible adenine nucleotide storage form in normal and ischaemic rat heart. Biosci Rep 5:7–12, 1985

    Google Scholar 

  12. Fitt PS, Korecky B, Sharma N: Further studies on the presumed adenine storage compound of rat heart. Biosci Rep 5:1061–1069, 1985

    Google Scholar 

  13. See YP, Fitt PS: A study of the localization of polynucleotide phosphorylase within rat liver cells and of its distribution among rat tissues and diverse animal species. Biochem J 130:355–362, 1972

    Google Scholar 

  14. Bates DJ, Perrett D, Mowbray J: Systematic variations in the content of purine nucleotides in the steady-state perfused rat heart. Biochem. J 176:485–493

  15. Gajda AT, de Behrens GZ, Fitt PS: Preparation, proteolysis and reversible oxidation of highly purified Azotobacter vinelandii polynucleotide phosphorylase. Biochem J 120:753–761, 1970

    Google Scholar 

  16. Randerath K, Randerath E: Thin-layer separation methods for nucleic acid derivatives. Meth Enzymol 12A:323–347, 1967

    Google Scholar 

  17. Palmer JM, Tandler B, Hoppel CL: Biochemical properties of subsarcolemmal and interfibrillar mitochondria isolated from rat cardiac muscle. J Biol Chem 252:8731–8739, 1977

    Google Scholar 

  18. Mela L, Seitz S: Isolation of mitochondria with emphasis on heart mitochondria from small amounts of tissue. Meth Enzymol 55:39–46, 1979

    Google Scholar 

  19. ordahl A, Johnson C, Blailock ZR, Schwartz A: The mitochondrion. Meth Parmacol 1:247–286, 1971

    Google Scholar 

  20. Layne E: Spectrophotometric and turbidimetric methods for measuring proteins. Meth Enzymol 3:447–454, 1957

    Google Scholar 

  21. Godefroy-Colburn T, Grunberg-Manago M: Polynucleotide phosphorylase. In: Boyer PD (ed) The Enzymes 3rd Ed. Academic Press, New York, 1972, pp 533–574

    Google Scholar 

  22. Perry RP, Kelley DE, Latorre J: Synthesis and turnover of nuclear and cytoplasmic polyadenylic acid in mouse L cells. J Mol Biol 82:315–331, 1974

    Google Scholar 

  23. Brawerman G: The role of the polyA sequences in mammalian messenger RNA. CRC Crit Rev. Biochem. 10:1–38, 1981

    Google Scholar 

  24. Attardi G: Organization and expression of the mammalian mitochondrial genome: a lesson in economy. TIBS 6:86–89, 1981

    Google Scholar 

  25. Clayton DA: Transcription of the mammalian mitochondrial genome. Ann Rev Biochem 53:573–594, 1984

    Google Scholar 

  26. Kroon AM, Saconne C, Botman MJ: RNA and protein synthesis by sterile rat liver mitochondria. Biochim Biophys Acta 142:552–554, 1967

    Google Scholar 

  27. Fukamachi S, Bartoov B, Freeman KB: Synthesis of ribonucleic acid by isolated rat liver mitochondria. Biochem J 128:299–309, 1972

    Google Scholar 

  28. Rose KM, Jacob ST: Polyadenylic acid synthesis in isolated rat liver mitochondria. Biochemistry 15:5046–5052, 1976

    Google Scholar 

  29. Kantharaj GR, Bhat KS, Avadhani NG: Mode of transcription and maturation of ribosomal ribonucleic acid in vitro in mitochondria from Ehrlich ascites cells. Biochemistry 22:3151–56, 1983

    Google Scholar 

  30. Winkler HH, Bygrave FL, Lehninger AL: Characterization of the atractyloside-sensitive adenine nucleotide transport system in rat liver mitochondria. J Biol Chem 243:20–28, 1968

    Google Scholar 

  31. Bukoski RD, Sparks HV, Mela LM: Rat heart mitochondria release adenosine. Biochem Biophys Res Commun 113:990–995, 1983

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fitt, P.S., Sharma, N., Attial, J. et al. Studies of adenosine incorporation in Langendorff rat heart and rat heart mitochondria. Mol Cell Biochem 78, 37–46 (1987). https://doi.org/10.1007/BF00224422

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00224422

Key words

Navigation