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. 1969 Apr 1;41(1):124–132. doi: 10.1083/jcb.41.1.124

PLASMA MEMBRANES OF THE RAT LIVER

Isolation and Enzymatic Characterization of a Fraction Rich in Bile Canaliculi

C S Song 1, W Rubin 1, A B Rifkind 1, A Kappas 1
PMCID: PMC2107735  PMID: 4304740

Abstract

A method is described for the rapid isolation of a plasma membrane fraction containing a high concentration of intact bile canaliculi from the rat liver. Isolated bile canaliculi retain most of the ultrastructural features exhibited in the intact liver cell. The final fraction contains 5'-nucleotidase activity at approximately the same concentration as that in previous preparations of plasma membranes. In the presence of 0.01 M Mg++, 5'-nucleotidase exhibits a double pH optimum at pH values of 7.5 and 9.5. The activities of glucose-6-phosphatase and alkaline phosphatase are present in low amounts. Cytochrome P-450 is not detectable. Na+-K+-activation of ATPase is observed to the extent of 20–36% in about half of the assays. The availability of a method for preparation of intact bile canaliculi should prove useful for studying the biochemical events associated with the transport of bile constituents into canaliculi.

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Selected References

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  1. BIAVA C. G. STUDIES ON CHOLESTASIS. A RE-EVALUATION OF THE FINE STRUCTURE OF NORMAL HUMAN BILE CANALICULI. Lab Invest. 1964 Aug;13:840–864. [PubMed] [Google Scholar]
  2. BODANSKY O., SCHWARTZ M. K. COMPARATIVE EFFECTS OF L-HISTIDINE ON THE ACTIVITIES OF 5'-NUCLEOTIDASE AND ALKALINE PHOSPHATASE. J Biol Chem. 1963 Oct;238:3420–3427. [PubMed] [Google Scholar]
  3. BONTING S. L., CARAVAGGIO L. L., HAWKINS N. M. Studies on sodium-potassium-activated adenosinetriphosphatase. IV. Correlation with cation transport sensitive to cardiac glycosides. Arch Biochem Biophys. 1962 Sep;98:413–419. doi: 10.1016/0003-9861(62)90206-0. [DOI] [PubMed] [Google Scholar]
  4. Barclay M., Barclay R. K., Essner E. S., Skipski V. P., Terebus-Kekish O. Plasma membranes of rat liver: isolation of lipoprotein macromolecules. Science. 1967 May 5;156(3775):665–667. doi: 10.1126/science.156.3775.665. [DOI] [PubMed] [Google Scholar]
  5. DAEMS W. T. The micro-anatomy of the smallestbiliary pathways in mouse liver tissue. Acta Anat (Basel) 1961;46:1–24. doi: 10.1159/000141765. [DOI] [PubMed] [Google Scholar]
  6. Dod B. J., Gray G. M. The lipid composition of rat-liver plasma membranes. Biochim Biophys Acta. 1968 Apr 29;150(3):397–404. doi: 10.1016/0005-2736(68)90138-7. [DOI] [PubMed] [Google Scholar]
  7. ELIAS H. A re-examination of the structure of the mammalian liver; the hepatic lobule and its relation to the vascular and biliary systems. Am J Anat. 1949 Nov;85(3):379-456, 15 pl. doi: 10.1002/aja.1000850303. [DOI] [PubMed] [Google Scholar]
  8. EMMELOT P., BOS C. J., BENEDETTI E. L., RUEMKE P. STUDIES ON PLASMA MEMBRANES. I. CHEMICAL COMPOSITION AND ENZYME CONTENT OF PLASMA MEMBRANES ISOLATED FROM RAT LIVER. Biochim Biophys Acta. 1964 Jul 15;90:126–145. doi: 10.1016/0304-4165(64)90125-4. [DOI] [PubMed] [Google Scholar]
  9. Emmelot P., Bos C. J. Studies on plasma membranes. 3. Mg2+-ATPase,(Na+-K+-Mg2+)-ATPase and 5'-nucleotidase activity of plasma membranes isolated from rat liver. Biochim Biophys Acta. 1966 Jul 13;120(3):369–382. doi: 10.1016/0926-6585(66)90304-9. [DOI] [PubMed] [Google Scholar]
  10. Emmelot P., Bos C. J. Studies on plasma membranes. VI. Differences in the effect of temperature on the ATPase and (Na+-K+)-ATPase activities of plasma membranes isolated from rat liver and hepatoma. Biochim Biophys Acta. 1968 Apr 29;150(3):354–363. doi: 10.1016/0005-2736(68)90134-x. [DOI] [PubMed] [Google Scholar]
  11. FARQUHAR M. G., PALADE G. E. Junctional complexes in various epithelia. J Cell Biol. 1963 May;17:375–412. doi: 10.1083/jcb.17.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. FAWCETT D. W. Observations on the cytology and electron microscopy of hepatic cells. J Natl Cancer Inst. 1955 Apr;15(5 Suppl):1475–1503. [PubMed] [Google Scholar]
  13. Gillette J. R. Biochemistry of drug oxidation and reduction by enzymes in hepatic endoplasmic reticulum. Adv Pharmacol. 1966;4:219–261. doi: 10.1016/s1054-3589(08)60100-3. [DOI] [PubMed] [Google Scholar]
  14. Heath T., Wissig S. L. Fine structure of the surface of mouse hepatic cells. Am J Anat. 1966 Jul;119(1):97–127. doi: 10.1002/aja.1001190107. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Levin S. J., Bodansky O. The double pH optimum of 5'-nucleotidase of bull seminal plasma. J Biol Chem. 1966 Jan 10;241(1):51–56. [PubMed] [Google Scholar]
  18. NEVILLE D. M., Jr The isolation of a cell membrane fraction from rat liver. J Biophys Biochem Cytol. 1960 Oct;8:413–422. doi: 10.1083/jcb.8.2.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. OMURA T., SATO R. THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE. J Biol Chem. 1964 Jul;239:2370–2378. [PubMed] [Google Scholar]
  20. PALADE G. E., SIEKEVITZ P. Liver microsomes; an integrated morphological and biochemical study. J Biophys Biochem Cytol. 1956 Mar 25;2(2):171–200. doi: 10.1083/jcb.2.2.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. POST R. L., MERRITT C. R., KINSOLVING C. R., ALBRIGHT C. D. Membrane adenosine triphosphatase as a participant in the active transport of sodium and potassium in the human erythrocyte. J Biol Chem. 1960 Jun;235:1796–1802. [PubMed] [Google Scholar]
  22. SKOU J. C. ENZYMATIC BASIS FOR ACTIVE TRANSPORT OF NA+ AND K+ ACROSS CELL MEMBRANE. Physiol Rev. 1965 Jul;45:596–617. doi: 10.1152/physrev.1965.45.3.596. [DOI] [PubMed] [Google Scholar]
  23. SKOU J. C. The influence of some cations on an adenosine triphosphatase from peripheral nerves. Biochim Biophys Acta. 1957 Feb;23(2):394–401. doi: 10.1016/0006-3002(57)90343-8. [DOI] [PubMed] [Google Scholar]
  24. Song C. S., Bodansky O. Subcellular localization and properties of 5'-nucleotidase in the rat liver. J Biol Chem. 1967 Feb 25;242(4):694–699. [PubMed] [Google Scholar]
  25. VENABLE J. H., COGGESHALL R. A SIMPLIFIED LEAD CITRATE STAIN FOR USE IN ELECTRON MICROSCOPY. J Cell Biol. 1965 May;25:407–408. doi: 10.1083/jcb.25.2.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. WOOD R. L. Some structural features of the bile canaliculus in calf liver. Anat Rec. 1961 Jul;140:207–215. doi: 10.1002/ar.1091400306. [DOI] [PubMed] [Google Scholar]

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