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. 1986 Jan;49(1):307–317. doi: 10.1016/s0006-3495(86)83643-8

Structure of the Channels in the Outer Mitochondrial Membrane

Electron Microscopic Studies of the Periodic Arrays Induced by Phospholipase a2 Treatment of the Neurospora membrane

C A Mannella, A Ribeiro, J Frank
PMCID: PMC1329639  PMID: 19431637

Abstract

The channel proteins in outer membranes of Neurospora crassa mitochondria spontaneously organize into periodic arrays when the membranes are dialyzed in the presence of soluble phospholipase A2. Electron microscopic images have been recorded at different electron doses from channel arrays in a variety of negative stains, as well as in vitreous ice. Fourier or correlation averages are formed from image fields containing a few hundred unit cells. These averages can be subsequently classified by correspondence analysis and summed to form representative averages over thousands of unit cells. In averages of negatively stained arrays, the stain-filled channel openings are bounded by smaller stain-excluding maxima. The projections of the channel openings are smaller and the subsidiary maxima are more pronounced for channel arrays contrasted with dilute (0.1%) uranyl acetate or aurothioglucose than for arrays embedded in 1% uranyl acetate. Projection images of unstained, ice-embedded membranes provide direct information about distribution of protein and lipid in the VDAC arrays. First experiments have yielded density maps with an apparent Fourier resolution of ∼1/(2 nm).

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Images in this article

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Selected References

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

  1. Brisson A., Unwin P. N. Quaternary structure of the acetylcholine receptor. Nature. 1985 Jun 6;315(6019):474–477. doi: 10.1038/315474a0. [DOI] [PubMed] [Google Scholar]
  2. Colombini M. A candidate for the permeability pathway of the outer mitochondrial membrane. Nature. 1979 Jun 14;279(5714):643–645. doi: 10.1038/279643a0. [DOI] [PubMed] [Google Scholar]
  3. Frank J., Goldfarb W., Eisenberg D., Baker T. S. Reconstruction of glutamine synthetase using computer averaging. Ultramicroscopy. 1978;3(3):283–290. doi: 10.1016/s0304-3991(78)80038-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Frank J., Verschoor A., Boublik M. Multivariate statistical analysis of ribosome electron micrographs. L and R lateral views of the 40 S subunit from HeLa cells. J Mol Biol. 1982 Oct 15;161(1):107–133. doi: 10.1016/0022-2836(82)90281-9. [DOI] [PubMed] [Google Scholar]
  5. Freitag H., Neupert W., Benz R. Purification and characterisation of a pore protein of the outer mitochondrial membrane from Neurospora crassa. Eur J Biochem. 1982 Apr;123(3):629–636. doi: 10.1111/j.1432-1033.1982.tb06578.x. [DOI] [PubMed] [Google Scholar]
  6. Hallermayer G., Neupert W. Lipid composition of mitochondrial outer and inner membranes of Neurospora crassa. Hoppe Seylers Z Physiol Chem. 1974 Mar;355(3):279–288. doi: 10.1515/bchm2.1974.355.1.279. [DOI] [PubMed] [Google Scholar]
  7. Kühlbrandt W. Discrimination of protein and nucleic acids by electron microscopy using contrast variation. Ultramicroscopy. 1982;7(3):221–232. doi: 10.1016/0304-3991(82)90169-3. [DOI] [PubMed] [Google Scholar]
  8. Makowski L., Caspar D. L., Phillips W. C., Goodenough D. A. Gap junction structures. V. Structural chemistry inferred from X-ray diffraction measurements on sucrose accessibility and trypsin susceptibility. J Mol Biol. 1984 Apr 15;174(3):449–481. doi: 10.1016/0022-2836(84)90331-0. [DOI] [PubMed] [Google Scholar]
  9. Mannella C. A., Bonner W. D., Jr X-ray diffraction from oriented outer mitochondrial membranes. Detection of in-plane subunit structure. Biochim Biophys Acta. 1975 Dec 1;413(2):226–233. doi: 10.1016/0005-2736(75)90106-6. [DOI] [PubMed] [Google Scholar]
  10. Mannella C. A., Colombini M. Evidence that the crystalline arrays in the outer membrane of Neurospora mitochondria are composed of the voltage-dependent channel protein. Biochim Biophys Acta. 1984 Jul 25;774(2):206–214. doi: 10.1016/0005-2736(84)90293-1. [DOI] [PubMed] [Google Scholar]
  11. Mannella C. A., Colombini M., Frank J. Structural and functional evidence for multiple channel complexes in the outer membrane of Neurospora crassa mitochondria. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2243–2247. doi: 10.1073/pnas.80.8.2243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mannella C. A., Frank J. Negative staining characteristics of arrays of mitochondrial pore protein: use of correspondence analysis to classify different staining patterns. Ultramicroscopy. 1984;13(1-2):93–102. doi: 10.1016/0304-3991(84)90060-3. [DOI] [PubMed] [Google Scholar]
  13. Mannella C. A. Phospholipase-induced crystallization of channels in mitochondrial outer membranes. Science. 1984 Apr 13;224(4645):165–166. doi: 10.1126/science.6322311. [DOI] [PubMed] [Google Scholar]
  14. Mannella C. A. Structure of the outer mitochondrial membrane: ordered arrays of porelike subunits in outer-membrane fractions from Neurospora crassa mitochondria. J Cell Biol. 1982 Sep;94(3):680–687. doi: 10.1083/jcb.94.3.680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Munn E. A. The application of the negative staining technique to the study of membranes. Methods Enzymol. 1974;32:20–35. doi: 10.1016/0076-6879(74)32005-8. [DOI] [PubMed] [Google Scholar]
  16. Nikaido H., Luckey M., Rosenberg E. Y. Nonspecific and specific diffusion channels in the outer membrane of Escherichia coli. J Supramol Struct. 1980;13(3):305–313. doi: 10.1002/jss.400130304. [DOI] [PubMed] [Google Scholar]
  17. Ross M. J., Klymkowsky M. W., Agard D. A., Stroud R. M. Structural studies of a membrane-bound acetylcholine receptor from Torpedo californica. J Mol Biol. 1977 Nov;116(4):635–659. doi: 10.1016/0022-2836(77)90264-9. [DOI] [PubMed] [Google Scholar]
  18. Schein S. J., Colombini M., Finkelstein A. Reconstitution in planar lipid bilayers of a voltage-dependent anion-selective channel obtained from paramecium mitochondria. J Membr Biol. 1976 Dec 28;30(2):99–120. doi: 10.1007/BF01869662. [DOI] [PubMed] [Google Scholar]
  19. Unwin P. N., Zampighi G. Structure of the junction between communicating cells. Nature. 1980 Feb 7;283(5747):545–549. doi: 10.1038/283545a0. [DOI] [PubMed] [Google Scholar]
  20. Zalman L. S., Nikaido H., Kagawa Y. Mitochondrial outer membrane contains a protein producing nonspecific diffusion channels. J Biol Chem. 1980 Mar 10;255(5):1771–1774. [PubMed] [Google Scholar]

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