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The Journal of Membrane Biology

, Volume 20, Issue 1, pp 99–110 | Cite as

Isolation and characterization of the plasma membrane from Yoshida hepatoma cells

  • A. Réthy
  • A. Trevisani
  • R. Manservigi
  • V. Tomasi
Article

Summary

Plasma membranes isolated from Yoshida ascites hepatoma AH-130 by a modification of the method of T. K. Ray (Biochim. Biophys. Acta196: 1, 1970), were subfractionated into three fractions having densities (d) 1.12, 1.14 and 1.16 by discontinuous sucrose density-gradient. Membrane subfractions were characterized by electron-microscopy, by assay of marker enzymes and by lipid composition. All subfractions appeared to be essentially free from whole mitochondria, lysosomes and nuclei. Subfraction d 1.16 had, the highest 5′-nucleotidase, Mg++-ATPase and (Na++K+)-ATPase activities; cytochromec oxidase was undetectable in any fraction and glucose-6-phosphatase was measurable only in fraction d 1.14. Adenylate cyclase had the highest activity in fractions d 1.14 and 1.16. Cyclic AMP phosphodiesterase was nearly equally distributed in the fractions. Adenylate, cyclase, 5′-nucleotidase and Mg++-ATPase activities of tumor membrane were lower with respect to liver plasma membrane, while cyclic AMP phosphodiesterase and (Na++K+)-ATPase were found to have similar activities in the two membrane preparations. With respect to liver membrane, hepatoma membrane contained a higher amount of glycolipids and a higher amount of phospholipids accounted for mainly, by sphingomyelin, phosphatidylserine and phosphatidic acid. The possible significance of the decrease of adenylate activity in the hepatoma membrane is briefly discussed.

Keywords

ATPase Activity Hepatoma Cell Adenylate Cyclase Lipid Composition Phosphatidylserine 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag New York Inc 1975

Authors and Affiliations

  • A. Réthy
    • 1
  • A. Trevisani
    • 1
  • R. Manservigi
    • 1
  • V. Tomasi
    • 1
  1. 1.Institute of General PhysiologyUniversity of FerraraFerraraItaly

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