Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1970 Jul 1;46(1):137–150. doi: 10.1083/jcb.46.1.137

THE SURFACE CHARGE OF RAT LIVER MITOCHONDRIA AND THEIR MEMBRANES

Clarification of Some Controversies Concerning Mitochondrial Structure

Hans-G Heidrich 1, Roland Stahn 1, Kurt Hannig 1
PMCID: PMC2108077  PMID: 4318844

Abstract

The surface charge of intact mitochondria and submitochondrial particles was examined by the technique of preparative free flow electrophoresis. When submitochondrial preparations obtained by a swelling-contraction procedure were examined with this technique, two fractions were observed. One of these fractions exhibited the same electrophoretic properties as intact mitochondria, which indicated that it was derived from the outer limiting membrane of the mitochondrion. This fraction was found to contain the enzymes monoamine oxidase and rotenone-insensitive NADH-cytochrome c reductase which have been reported to be localized in the outer mitochondrial membrane. The other fraction exhibited an electrophoretic mobility which was different from that of intact mitochondria, and this fraction contained enzymes characteristic of the inner membrane-matrix fraction such as soluble and particulate enzymes of the Krebs cycle. Microsomes exhibited an electrophoretic mobility which was almost identical with that of the outer mitochondrial membrane. In addition to resolving the localization of enzymes in mitochondrial membranes, these data indicate that the outer limiting membrane of the mitochondrion is the sole determinant of the surface charge of mitochondria.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. ARRIGONI O., SINGER T. P. Limitations of the phenazine methosulphate assay for succinic and related dehydrogenases. Nature. 1962 Mar 31;193:1256–1258. doi: 10.1038/1931256a0. [DOI] [PubMed] [Google Scholar]
  2. Allmann D. W., Bachmann E., Orme-Johnson N., Tan W. C., Green D. E. Membrane systems of mitochondria. VI. Membranes of liver mitochondria. Arch Biochem Biophys. 1968 Jun;125(3):981–1012. doi: 10.1016/0003-9861(68)90537-7. [DOI] [PubMed] [Google Scholar]
  3. BRAUNITZER G., HOBOM G., HANNIG K. EIN EINFACHES VERFAHREN ZUR SELEKTIONIERUNG VON MUTANTEN. Hoppe Seylers Z Physiol Chem. 1964;338:278–280. doi: 10.1515/bchm2.1964.338.1-2.278. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Beattie D. S. Enzyme localization in the inner and outer membranes of rat liver mitochondria. Biochem Biophys Res Commun. 1968 Jun 28;31(6):901–907. doi: 10.1016/0006-291x(68)90537-8. [DOI] [PubMed] [Google Scholar]
  6. COOPERSTEIN S. J., LAZAROW A. A microspectrophotometric method for the determination of cytochrome oxidase. J Biol Chem. 1951 Apr;189(2):665–670. [PubMed] [Google Scholar]
  7. 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]
  8. DE DUVE C., PRESSMAN B. C., GIANETTO R., WATTIAUX R., APPELMANS F. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J. 1955 Aug;60(4):604–617. doi: 10.1042/bj0600604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Green D. E., Allmann D. W., Harris R. A., Tan W. C. Enzyme localization in the inner and outer mitochondrial membranes. Biochem Biophys Res Commun. 1968 May 10;31(3):368–378. doi: 10.1016/0006-291x(68)90485-3. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Hannig K., Krüsmann W. F. Die Anwendung der trägerfreien kontinuierlichen Elektrophorese zur Auftrennung der weissen Blutzellen aus Humanblut. Hoppe Seylers Z Physiol Chem. 1968 Feb;349(2):161–170. [PubMed] [Google Scholar]
  12. Hannig K., Stahn R., Maier K. P. Isolierung von Zellorganellfraktionen aus Rattenleber mit Hilfe der trägerfreien kontinuierlichen Elektrophorese. Hoppe Seylers Z Physiol Chem. 1969 Jun;350(6):784–786. [PubMed] [Google Scholar]
  13. Hannig K., Zeiller K. Zur Auftrennung und Charakterisierung immunkompetenter Zellen mit Hilfe der trägerfreien Ablenkungselektrophorese. Hoppe Seylers Z Physiol Chem. 1969 Apr;350(4):467–472. [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lévy M., Toury R., André J. Séparation des membranes mitochondriales. Purification et caractérisation enzymatique de la membrane externe. Biochim Biophys Acta. 1967 Sep 9;135(4):599–613. doi: 10.1016/0005-2736(67)90092-2. [DOI] [PubMed] [Google Scholar]
  17. Neupert W., Brdiczka D., Bücher T. Incorporation of amino acids into the outer and inner membrane of isolated rat liver mitochondria. Biochem Biophys Res Commun. 1967 May 25;27(4):488–493. doi: 10.1016/s0006-291x(67)80012-3. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Schnaitman C., Erwin V. G., Greenawalt J. W. The submitochondrial localization of monoamine oxidase. An enzymatic marker for the outer membrane of rat liver mitochondria. J Cell Biol. 1967 Mar;32(3):719–735. doi: 10.1083/jcb.32.3.719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schnaitman C., Greenawalt J. W. Enzymatic properties of the inner and outer membranes of rat liver mitochondria. J Cell Biol. 1968 Jul;38(1):158–175. doi: 10.1083/jcb.38.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Thompson J. E., Coleman R., Finean J. B. Some biochemical and X-ray diffraction studies of mitochondrial outer membrane. Biochim Biophys Acta. 1967;135(5):1074–1078. doi: 10.1016/0005-2736(67)90081-8. [DOI] [PubMed] [Google Scholar]
  23. Tipton K. F. The sub-mitochondrial localization of monoamine oxidase in rat liver and brain. Biochim Biophys Acta. 1967;135(5):910–920. doi: 10.1016/0005-2736(67)90060-0. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES