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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1969 Feb;62(2):612–619. doi: 10.1073/pnas.62.2.612

A FLUORESCENCE PROBE OF ENERGY-DEPENDENT STRUCTURE CHANGES IN FRAGMENTED MEMBRANES*

A Azzi 1, B Chance 1, G K Radda 1,, C P Lee 1
PMCID: PMC277854  PMID: 4307717

Abstract

The reaction of the fluorochrome, 8-anilino-1-naphthalene-sulfonic acid (ANS), with fragmented membranes from beef heart mitochondria has been studied. ANS fluorescence is found to be enhanced 25-fold on binding to the membrane fragments in the absence of energy conservation, and this enhancement is increased to 35-fold in the membrane energized by substrate plux oxygen. The fluorescence of bound ANS depends upon the energy state of the membrane fragments, as indicated by the effects of ATP, substrates of the respiratory chain, oligomycin, and uncouplers. It is concluded that the changes of ANS fluorescence indicate structural changes of the mitochondrial membrane associated with energy conservation. The time course of energization is readily followed by ANS, and has a half-time of two seconds at 26°.

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Selected References

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

  1. CHANCE B., COHEN P., JOBSIS F., SCHOENER B. Intracellular oxidation-reduction states in vivo. Science. 1962 Aug 17;137(3529):499–508. doi: 10.1126/science.137.3529.499. [DOI] [PubMed] [Google Scholar]
  2. CHANCE B., HOLLUNGER G. Energy-linked reduction of mitochondrial pyridine nucleotide. Nature. 1960 Mar 5;185:666–672. doi: 10.1038/185666a0. [DOI] [PubMed] [Google Scholar]
  3. CHANCE B., PACKER L. Light-scattering and absorption effects caused by addition of adenosine diphosphate to rat-heart-muscle sarcosomes. Biochem J. 1958 Feb;68(2):295–297. doi: 10.1042/bj0680295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. CHANCE B., WILLIAMS G. R., HOLMES W. F., HIGGINS J. Respiratory enzymes in oxidative phosphorylation. V. A mechanism for oxidative phosphorylation. J Biol Chem. 1955 Nov;217(1):439–451. [PubMed] [Google Scholar]
  5. CHANCE B., WILLIAMS G. R. Respiratory enzymes in oxidative phosphorylation. III. The steady state. J Biol Chem. 1955 Nov;217(1):409–427. [PubMed] [Google Scholar]
  6. Chance B., Lee C. P., Mela L. Control and conservation of energy in the cytochrome chain. Fed Proc. 1967 Sep;26(5):1341–1354. [PubMed] [Google Scholar]
  7. Chance B., Mela L. Energy-linked changes of hydrogen ion concentration in submitochondrial particles. J Biol Chem. 1967 Mar 10;242(5):830–844. [PubMed] [Google Scholar]
  8. Chance B., Schoener B. High and low energy states of cytochromes. I. In mitochondria. J Biol Chem. 1966 Oct 25;241(20):4567–4573. [PubMed] [Google Scholar]
  9. Chance B. The reactivity of haemoproteins and cytochromes. Biochem J. 1967 Apr;103(1):1–18. doi: 10.1042/bj1030001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chen R. F. Removal of fatty acids from serum albumin by charcoal treatment. J Biol Chem. 1967 Jan 25;242(2):173–181. [PubMed] [Google Scholar]
  11. Daniel E., Weber G. Cooperative effects in binding by bovine serum albumin. I. The binding of 1-anilino-8-naphthalenesulfonate. Fluorimetric titrations. Biochemistry. 1966 Jun;5(6):1893–1900. doi: 10.1021/bi00870a016. [DOI] [PubMed] [Google Scholar]
  12. Dodd G. H., Radda G. K. Interaction of glutamate dehydrogenase with fluorescent dyes. Biochem Biophys Res Commun. 1967 May 25;27(4):500–504. doi: 10.1016/s0006-291x(67)80014-7. [DOI] [PubMed] [Google Scholar]
  13. Green D. E., Asai J., Harris R. A., Penniston J. T. Conformational basis of energy transformations in membrane systems. 3. Configurational changes in the mitochondrial inner membrane induced by changes in functional states. Arch Biochem Biophys. 1968 May;125(2):684–705. doi: 10.1016/0003-9861(68)90626-7. [DOI] [PubMed] [Google Scholar]
  14. Hackenbrock C. R. Ultrastructural bases for metabolically linked mechanical activity in mitochondria. II. Electron transport-linked ultrastructural transformations in mitochondria. J Cell Biol. 1968 May;37(2):345–369. doi: 10.1083/jcb.37.2.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kurup C. K., Sanadi D. R. Stoichiometry of uncoupler to respiratory assembly in oxidative phosphorylation. Arch Biochem Biophys. 1968 Aug;126(2):722–724. doi: 10.1016/0003-9861(68)90459-1. [DOI] [PubMed] [Google Scholar]
  16. LOW H., KRUEGER H., ZIEGLER D. M. On the reduction of externally added DPN by succinate in submitochondrial particles. Biochem Biophys Res Commun. 1961 Jun 28;5:231–237. doi: 10.1016/0006-291x(61)90116-4. [DOI] [PubMed] [Google Scholar]
  17. Mitchell P., Moyle J. Evidence discriminating between the chemical and the chemiosmotic mechanisms of electron transport phosphorylation. Nature. 1965 Dec 18;208(5016):1205–1206. doi: 10.1038/2081205a0. [DOI] [PubMed] [Google Scholar]
  18. PACKER L. Metabolic and structural states of mitochondria. I. Regulation by adenosine diphosphate. J Biol Chem. 1960 Jan;235:242–249. [PubMed] [Google Scholar]
  19. Racker E., Horstman L. L. Partial resolution of the enzymes catalyzing oxidative phosphorylation. 13. Structure and function of submitochondrial particles completely resolved with respect to coupling factor. J Biol Chem. 1967 May 25;242(10):2547–2551. [PubMed] [Google Scholar]
  20. Stryer L. Fluorescence spectroscopy of proteins. Science. 1968 Nov 1;162(3853):526–533. doi: 10.1126/science.162.3853.526. [DOI] [PubMed] [Google Scholar]
  21. Stryer L. The interaction of a naphthalene dye with apomyoglobin and apohemoglobin. A fluorescent probe of non-polar binding sites. J Mol Biol. 1965 Sep;13(2):482–495. doi: 10.1016/s0022-2836(65)80111-5. [DOI] [PubMed] [Google Scholar]
  22. WEBER G., YOUNG L. B. FRAGMENTATION OF BOVINE SERUM ALBUMIN BY PEPSIN. I. THE ORIGIN OF THE ACID EXPANSION OF THE ALBUMIN MOLECULE. J Biol Chem. 1964 May;239:1415–1423. [PubMed] [Google Scholar]

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