Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1985 Jun 1;228(2):493–503. doi: 10.1042/bj2280493

Kinetic studies of the uptake of aspartate aminotransferase and malate dehydrogenase into mitochondria in vitro.

E Marra, S Passarella, E Casamassima, E Perlino, S Doonan, E Quagliariello
PMCID: PMC1145008  PMID: 4015628

Abstract

Kinetic measurements of the uptake of native mitochondrial aspartate aminotransferase and malate dehydrogenase into mitochondria in vitro were carried out. The uptake of both the enzymes is essentially complete in 1 min and shows saturation characteristics. The rate of uptake of aspartate aminotransferase into mitochondria is decreased by malate dehydrogenase, and vice versa. The inhibition is exerted by isoenzyme remaining outside the mitochondria rather than by isoenzyme that has been imported. The thiol compound beta-mercaptoethanol decreases the rate of uptake of the tested enzymes; inhibition is a result of interaction of beta-mercaptoethanol with the mitochondria and not with the enzymes themselves. The rate of uptake of aspartate aminotransferase is inhibited non-competitively by malate dehydrogenase, but competitively by beta-mercaptoethanol. The rate of uptake of malate dehydrogenase is inhibited non-competitively by aspartate aminotransferase and by beta-mercaptoethanol. beta-Mercaptoethanol prevents the inhibition of the rate of uptake of malate dehydrogenase by aspartate aminotransferase. These results are interpreted in terms of a model system in which the two isoenzymes have separate but interacting binding sites within a receptor in the mitochondrial membrane system.

Full text

PDF
493

Selected References

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

  1. Chua N. H., Schmidt G. W. Transport of proteins into mitochondria and chloroplasts. J Cell Biol. 1979 Jun;81(3):461–483. doi: 10.1083/jcb.81.3.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Coty W. A., Pedersen P. L. Phosphate transport in rat liver mitochondria. Kinetics, inhibitor sensitivity, energy requirements, and labeled components. Mol Cell Biochem. 1975 Nov 14;9(2):109–124. doi: 10.1007/BF01732202. [DOI] [PubMed] [Google Scholar]
  3. Doonan S., Marra E., Passarella S., Saccone C., Quagliariello E. Transport of proteins into mitochondria. Int Rev Cytol. 1984;91:141–186. doi: 10.1016/s0074-7696(08)61316-9. [DOI] [PubMed] [Google Scholar]
  4. Fonyó A. SH-group reagents as tools in the study of mitochondrial anion transport. J Bioenerg Biomembr. 1978 Dec;10(5-6):171–194. doi: 10.1007/BF00743106. [DOI] [PubMed] [Google Scholar]
  5. Hennig B., Neupert W. Assembly of cytochrome c. Apocytochrome c is bound to specific sites on mitochondria before its conversion to holocytochrome c. Eur J Biochem. 1981 Dec;121(1):203–212. doi: 10.1111/j.1432-1033.1981.tb06450.x. [DOI] [PubMed] [Google Scholar]
  6. Klingenberg M. Kinetic study of the tricarboxylate carrier in rat liver mitochondria. Eur J Biochem. 1972 Apr 24;26(4):587–594. doi: 10.1111/j.1432-1033.1972.tb01801.x. [DOI] [PubMed] [Google Scholar]
  7. Marra E., Doonan S., Saccone C., Quagliariello E. Selective permeability of rat liver mitochondria to purified aspartate aminotransferases in vitro. Biochem J. 1977 Jun 15;164(3):685–691. doi: 10.1042/bj1640685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Marra E., Doonan S., Saccone C., Quagliariello E. Studies of the selective permeation of radioactively labelled aspartate aminotransferase isozymes into mitochondria in vitro. Eur J Biochem. 1978 Feb;83(2):427–435. doi: 10.1111/j.1432-1033.1978.tb12109.x. [DOI] [PubMed] [Google Scholar]
  9. Marra E., Passarella S., Doonan S., Quagliariello E., Saccone C. Protease resistance of aspartate aminotransferase imported in mitochondria. FEBS Lett. 1980 Dec 15;122(1):33–36. doi: 10.1016/0014-5793(80)80395-4. [DOI] [PubMed] [Google Scholar]
  10. Marra E., Passarella S., Doonan S., Saccone C., Quagliariello E. The effect of sulfhydryl group reagents on the permeation of mitochondrial aspartate aminotransferase into mitochondria in vitro. Arch Biochem Biophys. 1979 Jul;195(2):269–279. doi: 10.1016/0003-9861(79)90353-9. [DOI] [PubMed] [Google Scholar]
  11. Mori M., Morita T., Ikeda F., Amaya Y., Tatibana M., Cohen P. P. Synthesis, intracellular transport, and processing of the precursors for mitochondrial ornithine transcarbamylase and carbamoyl-phosphate synthetase I in isolated hepatocytes. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6056–6060. doi: 10.1073/pnas.78.10.6056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Palmieri F., Quagliariello E., Klingenberger M. Kinetics and specificity of the oxoglutarate carrier in rat-liver mitochondria. Eur J Biochem. 1972 Sep 25;29(3):408–416. doi: 10.1111/j.1432-1033.1972.tb02003.x. [DOI] [PubMed] [Google Scholar]
  13. Passarella S., Marra E., Doonan S., Languino L. R., Saccone C., Quagliariello E. Uptake of aspartate aminotransferase into mitochondria in vitro depends on the transmembrane pH gradient. Biochem J. 1982 Feb 15;202(2):353–362. doi: 10.1042/bj2020353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Passarella S., Marra E., Doonan S., Quagliariello E. Selective permeability of rat liver mitochondria to purified malate dehydrogenase isoenzymes in vitro. Biochem J. 1980 Nov 15;192(2):649–658. doi: 10.1042/bj1920649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Passarella S., Marra E., Doonan S., Quagliariello E. Uptake of malate dehydrogenase into mitochondria in vitro. Some characteristics of the process. Biochem J. 1983 Jan 15;210(1):207–214. doi: 10.1042/bj2100207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Quagliariello E., Palmieri F., Prezioso G., Klingenberg M. Kinetics of succinate uptake by rat-liver mitochondria. FEBS Lett. 1969 Aug;4(4):251–254. doi: 10.1016/0014-5793(69)80247-4. [DOI] [PubMed] [Google Scholar]
  17. Reid G. A., Yonetani T., Schatz G. Import of proteins into mitochondria. Import and maturation of the mitochondrial intermembrane space enzymes cytochrome b2 and cytochrome c peroxidase in intact yeast cells. J Biol Chem. 1982 Nov 10;257(21):13068–13074. [PubMed] [Google Scholar]
  18. Schatz G., Butow R. A. How are proteins imported into mitochondria? Cell. 1983 Feb;32(2):316–318. doi: 10.1016/0092-8674(83)90450-6. [DOI] [PubMed] [Google Scholar]
  19. WADDELL W. J. A simple ultraviolet spectrophotometric method for the determination of protein. J Lab Clin Med. 1956 Aug;48(2):311–314. [PubMed] [Google Scholar]
  20. Zimmermann R., Neupert W. Transport of proteins into mitochondria. Posttranslational transfer of ADP/ATP carrier into mitochondria in vitro. Eur J Biochem. 1980 Aug;109(1):217–229. doi: 10.1111/j.1432-1033.1980.tb04787.x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES