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. 1976 Mar 1;68(3):724–739. doi: 10.1083/jcb.68.3.724

Identification and transmembranous localization of active cytochrome oxidase in reconstituted membranes of purified phospholipids by electron microscopy

PMCID: PMC2109659  PMID: 192731

Abstract

Cytochrome oxidase vesicles with high oxidase activity and respiratory control ratio (greater than 3.5) were characterized by the freeze-etch technique for electron microscopy. By the use of this technique, cytochrome oxidase is shown to be an inner membrane particle. By locating cross-fractured vesicles in the same preparation, cytochrome oxidase particles are shown to extend across the phospholipid bilayer membranes. When cytochrome oxidase is added to preformed liposomes respiratory control is not observed, but high oxidase activity is maintained. In this preparation the cytochrome oxidase particles are located on the outer vesicle membrane surface. These observations provide direct evidence that cytochrome oxidase is found in a transmembranous position in closed, activecytochrome oxidase vesicles having respiratory control.

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Selected References

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  1. Branton D. Fracture faces of frozen membranes. Proc Natl Acad Sci U S A. 1966 May;55(5):1048–1056. doi: 10.1073/pnas.55.5.1048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Capaldi R. A. A dynamic model of cell membranes. Sci Am. 1974 Mar;230(3):26–33. doi: 10.1038/scientificamerican0374-26. [DOI] [PubMed] [Google Scholar]
  3. HALL C. E. Measurement of globular protein molecules by electron microscopy. J Biophys Biochem Cytol. 1960 Jul;7:613–618. doi: 10.1083/jcb.7.4.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hinkle P. C., Kim J. J., Racker E. Ion transport and respiratory control in vesicles formed from cytochrome oxidase and phospholipids. J Biol Chem. 1972 Feb 25;247(4):1338–1339. [PubMed] [Google Scholar]
  5. Kagawa Y., Kandrach A., Racker E. Partial resolution of the enzymes catalyzing oxidative phosphorylation. XXVI. Specificity of phospholipids required for energy transfer reactions. J Biol Chem. 1973 Jan 25;248(2):676–684. [PubMed] [Google Scholar]
  6. Kuboyama M., Yong F. C., King T. E. Studies on cytochrome oxidase. 8. Preparation and some properties of cardiac cytochrome oxidase. J Biol Chem. 1972 Oct 25;247(20):6375–6383. [PubMed] [Google Scholar]
  7. Misra D. N., Das Gupta N. N. Distortion in dimensions produced by shadowing for electron microscopy. J R Microsc Soc. 1965 Sep;84(3):373–384. [PubMed] [Google Scholar]
  8. Pinto da Silva P., Branton D. Membrane splitting in freeze-ethching. Covalently bound ferritin as a membrane marker. J Cell Biol. 1970 Jun;45(3):598–605. doi: 10.1083/jcb.45.3.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Racker E. Reconstitution of cytochrome oxidase vesicles and conferral of sensitivity to energy transfer inhibitors. J Membr Biol. 1972 Dec 29;10(3):221–235. doi: 10.1007/BF01867856. [DOI] [PubMed] [Google Scholar]
  11. Schneider D. L., Kagawa Y., Racker E. Chemical modification of the inner mitochondrial membrane. J Biol Chem. 1972 Jun 25;247(12):4074–4079. [PubMed] [Google Scholar]
  12. Telford J. N., Matsumura F. Dieldrin binding in subcellular nerve components of cockroaches. An electron microscopic and autoradiographic study. J Econ Entomol. 1970 Jun;63(3):795–800. doi: 10.1093/jee/63.3.795. [DOI] [PubMed] [Google Scholar]
  13. Tillack T. W., Marchesi V. T. Demonstration of the outer surface of freeze-etched red blood cell membranes. J Cell Biol. 1970 Jun;45(3):649–653. doi: 10.1083/jcb.45.3.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Vail W. J., Riley R. K. The structure of cytochrome oxidase membranes. FEBS Lett. 1974 Apr 1;40(2):269–273. doi: 10.1016/0014-5793(74)80242-5. [DOI] [PubMed] [Google Scholar]

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