Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Apr 1;98(4):1497–1504. doi: 10.1083/jcb.98.4.1497

A domain-specific marker for the hepatocyte plasma membrane. III. Isolation of bile canalicular membrane by immunoadsorption

PMCID: PMC2113235  PMID: 6371022

Abstract

Previous immunolabeling studies (Roman, L.M., and A.L. Hubbard, 1983, J. Cell Biol., 96:1548-1558; Roman, L.M., and A.L. Hubbard, 1984, J. Cell Biol., 98:1488-1496, companion paper) established leucine aminopeptidase (LAP) as a specific marker for the bile canalicular (BC) domain of the rat hepatocyte plasma membrane (PM). In this study, we have isolated membrane from a sonicated PM vesicle fraction using anti- LAP-coated Staphylococcus aureus cells as a solid-phase immunoadsorbent. The extent and specificity of the immunoadsorption were assessed by following the behavior of LAP (the BC marker) and 32P- labeled membrane phospholipids (a uniform membrane marker). The BC fraction obtained was significantly enriched in LAP (yield: greater than 70% of PM-LAP). Alkaline phosphatase, 5'-nucleotidase, and a 110,000-dalton glycoprotein, HA-4, were enriched in the BC fraction to the same extent as LAP (enzyme or antigen/LAP = 1.0). However, alkaline phosphodiesterase I was not enriched to the same degree (enzyme/LAP = 0.5). Contamination of this BC fraction by membrane derived from the sinusoidal domain and endoplasmic reticulum, as determined from the distribution of the asialoglycoprotein receptor and NADH cytochrome c reductase, respectively, was small (less than 13%).

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Avruch J., Wallach D. F. Preparation and properties of plasma membrane and endoplasmic reticulum fragments from isolated rat fat cells. Biochim Biophys Acta. 1971 Apr 13;233(2):334–347. doi: 10.1016/0005-2736(71)90331-2. [DOI] [PubMed] [Google Scholar]
  2. Blitzer B. L., Boyer J. L. Cytochemical localization of Na+, K+-ATPase in the rat hepatocyte. J Clin Invest. 1978 Nov;62(5):1104–1108. doi: 10.1172/JCI109216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyer J. L., Allen R. M., Ng O. C. Biochemical separation of Na+,K+-ATPase from a "purified" light density, "canalicular"-enriched plasma membrane fraction from rat liver. Hepatology. 1983 Jan-Feb;3(1):18–28. doi: 10.1002/hep.1840030103. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Evans W. H. Fractionation of liver plasma membranes prepared by zonal centrifugation. Biochem J. 1970 Mar;116(5):833–842. doi: 10.1042/bj1160833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hubbard A. L., Ma A. Isolation of rat hepatocyte plasma membranes. II. Identification of membrane-associated cytoskeletal proteins. J Cell Biol. 1983 Jan;96(1):230–239. doi: 10.1083/jcb.96.1.230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hubbard A. L., Wall D. A., Ma A. Isolation of rat hepatocyte plasma membranes. I. Presence of the three major domains. J Cell Biol. 1983 Jan;96(1):217–229. doi: 10.1083/jcb.96.1.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hubbard A. L., Wilson G., Ashwell G., Stukenbrok H. An electron microscope autoradiographic study of the carbohydrate recognition systems in rat liver. I. Distribution of 125I-ligands among the liver cell types. J Cell Biol. 1979 Oct;83(1):47–64. doi: 10.1083/jcb.83.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Inoue M., Kinne R., Tran T., Arias I. M. Taurocholate transport by rat liver sinusoidal membrane vesicles: evidence of sodium cotransport. Hepatology. 1982 Sep-Oct;2(5):572–579. doi: 10.1002/hep.1840020510. [DOI] [PubMed] [Google Scholar]
  11. Inoue M., Kinne R., Tran T., Biempica L., Arias I. M. Rat liver canalicular membrane vesicles. Isolation and topological characterization. J Biol Chem. 1983 Apr 25;258(8):5183–5188. [PubMed] [Google Scholar]
  12. Ito A., Palade G. E. Presence of NADPH-cytochrome P-450 reductase in rat liver Golgi membranes. Evidence obtained by immunoadsorption method. J Cell Biol. 1978 Nov;79(2 Pt 1):590–597. doi: 10.1083/jcb.79.2.590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kawajiri K., Ito A., Omura T. Subfractionation of rat liver microsomes by immunoprecipitation and immunoadsorption methods. J Biochem. 1977 Mar;81(3):779–789. doi: 10.1093/oxfordjournals.jbchem.a131516. [DOI] [PubMed] [Google Scholar]
  14. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  15. Merisko E. M., Farquhar M. G., Palade G. E. Coated vesicle isolation by immunoadsorption on Staphylococcus aureus cells. J Cell Biol. 1982 Mar;92(3):846–857. doi: 10.1083/jcb.92.3.846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miljanich G. P., Brasier A. R., Kelly R. B. Partial purification of presynaptic plasma membrane by immunoadsorption. J Cell Biol. 1982 Jul;94(1):88–96. doi: 10.1083/jcb.94.1.88. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ray T. K. A modified method for the isolation of the plasma membrane from rat liver. Biochim Biophys Acta. 1970 Jan 6;196(1):1–9. doi: 10.1016/0005-2736(70)90159-8. [DOI] [PubMed] [Google Scholar]
  18. Roman L. M., Hubbard A. L. A domain-specific marker for the hepatocyte plasma membrane. II. Ultrastructural localization of leucine aminopeptidase to the bile canalicular domain of isolated rat liver plasma membranes. J Cell Biol. 1984 Apr;98(4):1488–1496. doi: 10.1083/jcb.98.4.1488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roman L. M., Hubbard A. L. A domain-specific marker for the hepatocyte plasma membrane: localization of leucine aminopeptidase to the bile canalicular domain. J Cell Biol. 1983 Jun;96(6):1548–1558. doi: 10.1083/jcb.96.6.1548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Scharschmidt B. F., Keeffe E. B. Isolation of a rat liver plasma membrane fraction of probable canalicular origin. Preparative technique, enzymatic profile, composition, and solute transport. Biochim Biophys Acta. 1981 Sep 7;646(3):369–381. doi: 10.1016/0005-2736(81)90305-9. [DOI] [PubMed] [Google Scholar]
  21. Skipski V. P., Barclay M., Archibald F. M., Terebus-Kekish O., Reichman E. S., Good J. J. Lipid composition of rat liver cell membranes. Life Sci. 1965 Sep;4(17):1673–1680. doi: 10.1016/0024-3205(65)90213-4. [DOI] [PubMed] [Google Scholar]
  22. Smith G. D., Peters T. J. Analytical subcellular fractionation of rat liver with special reference to the localisation of putative plasma membrane marker enzymes. Eur J Biochem. 1980 Feb;104(1):305–311. doi: 10.1111/j.1432-1033.1980.tb04429.x. [DOI] [PubMed] [Google Scholar]
  23. Takeuchi M., Terayama H. Preparation and chemical composition of rat liver cell membranes. Exp Cell Res. 1965 Oct;40(1):32–44. doi: 10.1016/0014-4827(65)90287-9. [DOI] [PubMed] [Google Scholar]
  24. Touster O., Aronson N. N., Jr, Dulaney J. T., Hendrickson H. Isolation of rat liver plasma membranes. Use of nucleotide pyrophosphatase and phosphodiesterase I as marker enzymes. J Cell Biol. 1970 Dec;47(3):604–618. doi: 10.1083/jcb.47.3.604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wall D. A., Hubbard A. L. Galactose-specific recognition system of mammalian liver: receptor distribution on the hepatocyte cell surface. J Cell Biol. 1981 Sep;90(3):687–696. doi: 10.1083/jcb.90.3.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wisher M. H., Evans W. H. Functional polarity of the rat hepatocyte surface membrane. Isolation and characterization of plasma-membrane subfractions from the blood-sinusoidal, bile-Canalicular and contiguous surfaces of the hepatocyte. Biochem J. 1975 Feb;146(2):375–388. doi: 10.1042/bj1460375. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES