Home LiteratureArticle Details
PMID: 6371022 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

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

The Journal of cell biology ·Vol. 98 ·No. 4 ·1984-04-00 ·Pages 1497-504

Roman LM, Hubbard AL

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%).

MeSH Terms
Animals Antibodies Antigen-Antibody Complex Cell Fractionation Cell Membrane/ultrastructure Fluorescent Antibody Technique Hydrogen-Ion Concentration Liver/ultrastructure Male Membrane Proteins/isolation & purification Microscopy, Electron Molecular Weight Phosphorus Radioisotopes Rats Rats, Inbred Strains Staphylococcus aureus/immunology Ultrasonics
Chemicals
Antibodies Antigen-Antibody Complex Membrane Proteins Phosphorus Radioisotopes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Roman L M
Hubbard A L
References (26)
26 references, click to expand
  1. 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):369-81 PMID: 6116503
  2. Galactose-specific recognition system of mammalian liver: receptor distribution on the hepatocyte cell surface.
    J Cell Biol. 1981 Sep;90(3):687-96 PMID: 6270158
  3. Coated vesicle isolation by immunoadsorption on Staphylococcus aureus cells.
    J Cell Biol. 1982 Mar;92(3):846-57 PMID: 7045138
  4. Taurocholate transport by rat liver sinusoidal membrane vesicles: evidence of sodium cotransport.
    Hepatology. 1982 Sep-Oct;2(5):572-9 PMID: 7118069
  5. Partial purification of presynaptic plasma membrane by immunoadsorption.
    J Cell Biol. 1982 Jul;94(1):88-96 PMID: 6749869
  6. 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 PMID: 6130040
  7. Isolation of rat hepatocyte plasma membranes. I. Presence of the three major domains.
    J Cell Biol. 1983 Jan;96(1):217-29 PMID: 6298249
  8. Isolation of rat hepatocyte plasma membranes. II. Identification of membrane-associated cytoskeletal proteins.
    J Cell Biol. 1983 Jan;96(1):230-9 PMID: 6681819
  9. Rat liver canalicular membrane vesicles. Isolation and topological characterization.
    J Biol Chem. 1983 Apr 25;258(8):5183-8 PMID: 6833295
  10. 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):1548-58 PMID: 6304108
  11. 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):1488-96 PMID: 6201493
  12. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY.
    Biochem J. 1963 Oct;89:114-23 PMID: 14097352
  13. Preparation and chemical composition of rat liver cell membranes.
    Exp Cell Res. 1965 Oct;40(1):32-44 PMID: 5320251
  14. Lipid composition of rat liver cell membranes.
    Life Sci. 1965 Sep;4(17):1673-80 PMID: 5863374
  15. A modified method for the isolation of the plasma membrane from rat liver.
    Biochim Biophys Acta. 1970 Jan 6;196(1):1-9 PMID: 4312696
  16. Fractionation of liver plasma membranes prepared by zonal centrifugation.
    Biochem J. 1970 Mar;116(5):833-42 PMID: 4315049
  17. Isolation of rat liver plasma membranes. Use of nucleotide pyrophosphatase and phosphodiesterase I as marker enzymes.
    J Cell Biol. 1970 Dec;47(3):604-18 PMID: 5497542
  18. Preparation and properties of plasma membrane and endoplasmic reticulum fragments from isolated rat fat cells.
    Biochim Biophys Acta. 1971 Apr 13;233(2):334-47 PMID: 4326970
  19. 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):375-88 PMID: 125584
  20. 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):1617-24 PMID: 1102604
  21. 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;72:248-54 PMID: 942051
  22. Subfractionation of rat liver microsomes by immunoprecipitation and immunoadsorption methods.
    J Biochem. 1977 Mar;81(3):779-89 PMID: 405382
  23. Cytochemical localization of Na+, K+-ATPase in the rat hepatocyte.
    J Clin Invest. 1978 Nov;62(5):1104-8 PMID: 213446
  24. 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):590-7 PMID: 214451
  25. 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 PMID: 511941
  26. 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):305-11 PMID: 7371635
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.98.4.1497
Published
1984-04-00
Pages
1497-504
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113235
Subset
IM
Grants
NIGMS NIH HHS · GM29185 · United States
External Links
PubMed source DOI: 10.1083/jcb.98.4.1497 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2