Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1968 Feb;106(3):711–717. doi: 10.1042/bj1060711

Structural changes in mitochondria induced by uncoupling reagents. The response to proteolytic enzymes

Eugene C Weinbach 1,2, Joel Garbus 1,2
PMCID: PMC1198563  PMID: 4170712

Abstract

Interaction of uncoupling reagents with bovine serum albumin markedly inhibited its hydrolysis by proteolytic enzymes. The inhibition presumably is due to conformational transitions in the protein substrate induced by the binding of the ligand–uncoupling reagents. The proteolysis of casein, a protein that does not bind these reagents, was not affected, indicating that the proteinases themselves were not inactivated. In contrast, interaction of uncoupling reagents with freshly isolated rat liver mitochondria enhanced their susceptibility to proteolytic enzymes. This was shown by an increase in the release of ninhydrin-reacting material, by an increase in free acid groups and by a decrease in the turbidity of the mitochondrial suspensions. These effects, although opposite in direction to those obtained with albumin, are also presumed to indicate structural changes in the mitochondrial proteins and a disorganization of the protein–phospholipid complex. It is suggested that such structural alterations are expressed functionally as the uncoupling of oxidative phosphorylation.

Full text

PDF
711

Selected References

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

  1. Allmann D. W., Bachmann E., Green D. E. The membrane systems of the mitochondrion. II. The K fraction of the outer membrane of beef heart mitochondria. Arch Biochem Biophys. 1966 Jul;115(1):165–171. doi: 10.1016/s0003-9861(66)81052-4. [DOI] [PubMed] [Google Scholar]
  2. Bachmann E., Allmann D. W., Green D. E. The membrane systems of the mitochondrion. I. The S fraction of the outer membrane of beef heart mitochondria. Arch Biochem Biophys. 1966 Jul;115(1):153–164. doi: 10.1016/s0003-9861(66)81051-2. [DOI] [PubMed] [Google Scholar]
  3. Beechey R. B. The uncoupling of respiratory-chain phosphorylation by 4,5,6,7-tetrachloro-2-trifluoromethylbenzimidazole. Biochem J. 1966 Jan;98(1):284–289. doi: 10.1042/bj0980284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Caplan A. I., Greenawalt J. W. Biochemical and ultrastructural properties of osmotically lysed rat-liver mitochondria. J Cell Biol. 1966 Dec;31(3):455–472. doi: 10.1083/jcb.31.3.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DIANZANI M. U. Action of papain and of trypsin on the morphology and some enzymatic activities of isolated mitochondria. Experientia. 1953 Sep;9(9):343–345. doi: 10.1007/BF02155841. [DOI] [PubMed] [Google Scholar]
  6. GOODMAN D. S. Preparation of human serum albumin free of long-chain fatty acids. Science. 1957 Jun 28;125(3261):1296–1297. doi: 10.1126/science.125.3261.1296. [DOI] [PubMed] [Google Scholar]
  7. Green D. E., Bachmann E., Allmann D. W., Perdue J. F. The membrane systems of the mitochondrion. III. The isolation and properties of the outer membrane of beef heart mitochondria. Arch Biochem Biophys. 1966 Jul;115(1):172–180. doi: 10.1016/s0003-9861(66)81053-6. [DOI] [PubMed] [Google Scholar]
  8. Green D. E., Tzagoloff A. The mitochondrial electron transfer chain. Arch Biochem Biophys. 1966 Sep 26;116(1):293–304. doi: 10.1016/0003-9861(66)90036-1. [DOI] [PubMed] [Google Scholar]
  9. HEYTLER P. G. uncoupling of oxidative phosphorylation by carbonyl cyanide phenylhydrazones. I. Some characteristics of m-Cl-CCP action on mitochondria and chloroplasts. Biochemistry. 1963 Mar-Apr;2:357–361. doi: 10.1021/bi00902a031. [DOI] [PubMed] [Google Scholar]
  10. Hackenbrock C. R. Ultrastructural bases for metabolically linked mechanical activity in mitochondria. I. Reversible ultrastructural changes with change in metabolic steady state in isolated liver mitochondria. J Cell Biol. 1966 Aug;30(2):269–297. doi: 10.1083/jcb.30.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LEHNINGER A. L. Water uptake and extrusion by mitochondria in relation to oxidative phosphorylation. Physiol Rev. 1962 Jul;42:467–517. doi: 10.1152/physrev.1962.42.3.467. [DOI] [PubMed] [Google Scholar]
  12. Parsons D. F. Recent advances correlating structure and function in mitochondria. Int Rev Exp Pathol. 1965;4:1–54. [PubMed] [Google Scholar]
  13. Parsons D. F., Williams G. R., Chance B. Characteristics of isolated and purified preparations of the outer and inner membranes of mitochondria. Ann N Y Acad Sci. 1966 Jul 14;137(2):643–666. doi: 10.1111/j.1749-6632.1966.tb50188.x. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. WEINBACH E. C. A procedure for isolating stable mitochondria from rat liver and kidney. Anal Biochem. 1961 Aug;2:335–343. doi: 10.1016/0003-2697(61)90006-9. [DOI] [PubMed] [Google Scholar]
  16. WEINBACH E. C., GARBUS J. PROTEIN AS THE MITOCHONDRIAL SITE FOR ACTION OF UNCOUPLING PHENOLS. Science. 1964 Aug 21;145(3634):824–826. doi: 10.1126/science.145.3634.824. [DOI] [PubMed] [Google Scholar]
  17. WEINBACH E. C., GARBUS J. THE INTERACTION OF UNCOUPLING PHENOLS WITH MITOCHONDRIA AND WITH MITOCHONDRIAL PROTEIN. J Biol Chem. 1965 Apr;240:1811–1819. [PubMed] [Google Scholar]
  18. WEINBACH E. C. Oxidative phosphorylation with endogenous substrates of mitochondria. J Biol Chem. 1961 May;236:1526–1530. [PubMed] [Google Scholar]
  19. WEINBACH E. C. The effect of pentachlorophenol on oxidative phosphorylation. J Biol Chem. 1954 Oct;210(2):545–550. [PubMed] [Google Scholar]
  20. Weinbach E. C., Garbus J. Restoration by albumin of oxidative phosphorylation and related reactions. J Biol Chem. 1966 Jan 10;241(1):169–175. [PubMed] [Google Scholar]
  21. Weinbach E. C., Garbus J. The rapid restoration of respiratory control to uncoupled mitochondria. J Biol Chem. 1966 Aug 25;241(16):3708–3713. [PubMed] [Google Scholar]

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

RESOURCES