Two-Phase Partitioning of Rat Liver Plasma Membranes

  • Peter Gierow
  • Bengt Jergil
  • Per-Åke Albertsson


Rat liver plasma membranes were subjected to countercurrent distribution in a dextran/poly(ethylene glycol) two-phase system. The lateral heterogeneity was examined by plotting the specific activities of marker components, for each distribution fraction, against each other. The patterns obtained were compared with those of different model membranes. By this method 5′-nucleotidase and leucine amino-peptidase were found in separate but adjacent domains, whereas asialo orosomucoid receptors resided in a domain separated from those containing 5′-nucleotidase or leucine aminopeptidase by a domain lacking these markers.


Fragment Size Leucine Aminopeptidase Lateral Heterogeneity Countercurrent Distribution Hepatocyte Plasma Membrane 


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  1. 1.
    D.M. Neville, Jr., The isolation of a cell membrane fraction from rat liver, J. Biochem. Biophys. Cytol. 8:413 (1960)CrossRefGoogle Scholar
  2. 2.
    N.N. Aronson and O. Touster, Isolation of rat liver plasma membrane fragments in isotonic sucrose, Methods Enzymol. 31:90 (1974)PubMedCrossRefGoogle Scholar
  3. 3.
    M.H. Wisher and W.H. Evans, Functional polarity of the rat hepatocyte surface membrane. Isolation and characterisation of plasma-membrane subfractions from the blood-sinusoidal, bile canalicular and contiguous surfaces of the hepatocyte, Biochem. J. 146:375 (1975)PubMedGoogle Scholar
  4. 4.
    A.L. Hubbard, D.A. Wall and A. Ma, Isolation of rat hepatocyte plasma membranes. 1. Presence of the three major domains, J. Cell Biol. 96:217 (1983)PubMedCrossRefGoogle Scholar
  5. 5.
    J.A. Taylor, D. Lawson and J.D Judah, Analysis and purification of the blood sinusoidal domain of rat liver plasma membrane, Biochim. Biophys. Acta 732:154 (1983)PubMedCrossRefGoogle Scholar
  6. 6.
    P.J. Meier, E.S. Sztul, A. Reuben and J.L. Boyer, Structural and functional polarity of canalicular and basolateral plasma membrane vesicles isolated in high yield from rat liver, J. Cell Biol 98:991 (1984)PubMedCrossRefGoogle Scholar
  7. 7.
    P. Gierow, M. Sommarin, C. Larsson and B. Jergil, Fractionation of rat liver plasma-membrane regions by two-phase partitioning, Biochem. J. 235:685 (1986)PubMedGoogle Scholar
  8. 8.
    P.-Å. Albertsson, Analysis of the domain structure of membranes by fragmentation and separation in aqueous polymer two-phase systems, Quart. Rev. Biophys. 21:61 (1988)CrossRefGoogle Scholar
  9. 9.
    M.J. López-Pérez, G. Paris and C. Larsson, Highly purified mitochondria from rat brain prepared by phase partition, Biochim. Biophys. Acta 635:359 (1981)PubMedCrossRefGoogle Scholar
  10. 10.
    P. Gierow, B. Jergil and P.-Å. Albertsson, Lateral heterogeneity of rat liver plasma membranes analysed by countercurrent distribution, Biochem. J. 249:369 (1988)PubMedGoogle Scholar
  11. 11.
    H.-E. Åkerlund, An apparatus for countercurrent distribution in a centrifugal acceleration field, J. Biochem. Biophys. Methods 9:133 (1984)PubMedCrossRefGoogle Scholar
  12. 12.
    O.H. Lowry, N.J. Rosebrough, A.L. Farr and R.J. Randall, Protein measurement with the folin phenol reagent, J. Biol. Chem. 193:265 (1951)PubMedGoogle Scholar
  13. 13.
    J. Avruch and D.F.H. Wallach, Preparation and properties of plasma membrane and endoplasmic reticulum fragments from rat liver cells, Biochim. Biophys. Acta 233:334 (1971)PubMedCrossRefGoogle Scholar
  14. 14.
    S. Matsuura, H. Nakada, T. Sawamura and Y. Tashiro, Distribution of an asialoglycoprotein receptor on rat hepatocyte cell surface, J. Cell Biol. 95:864 (1982)PubMedCrossRefGoogle Scholar
  15. 15.
    L.M. Roman and A.L. Hubbard, A domain-specific marker for the hepatocyte plasma membrane. Localisation of leucine aminopeptidase to the bile canalicular domain, J. Cell Biol. 96:1548 (1983)PubMedCrossRefGoogle Scholar
  16. 16.
    S. Matsuura, S. Eto, K. Kato and Y. Tashiro, Ferritin immunoelectron microscopic localisation of 5′-nucloetidase on rat liver cell surface, J. Cell Biol. 99:166 (1984)PubMedCrossRefGoogle Scholar
  17. 17.
    J.R. Bartles, L.T. Braiterman and A.L. Hubbard, Endogenous and exogenous domain markers of the rat hepatocyte membrane, J. Cell Biol. 100:1126 (1985)PubMedCrossRefGoogle Scholar
  18. 18.
    C.W. Slife, M.D. Dorsett, G.T. Bouquett, A. Register, E. Taylor and S. Conroy, Subcellular localisation of a membrane-associated transglutaminase activity in rat liver, Arch. Biochem. Biophys. 241:329 (1985)PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Peter Gierow
    • 1
  • Bengt Jergil
    • 1
  • Per-Åke Albertsson
    • 1
  1. 1.Department of Biochemistry Chemical CentreUniversity of LundLundSweden

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