Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Mar;78(3):1557–1561. doi: 10.1073/pnas.78.3.1557

Immunological evidence for the transmembrane nature of the rat liver receptor for asialoglycoproteins.

J Harford, G G Ashwell
PMCID: PMC319170  PMID: 6262814

Abstract

Antibodies raised in goats against the rat hepatic receptor for desialylated glycoproteins were perfused through a rat liver and were specifically retained by the liver. These antireceptor antibodies also bound specifically to hepatocyte plasma membranes oriented with their cytoplasmic surface outward on polylysine-derivatized beads. These two phenomena were judged to be properties of distinct subpopulations of the antibody preparation because: (i) maximal adsorption of antibodies with membranes on polylysine beads did not affect subsequent retention by the perfused liver, and (ii) whereas perfusion resulted in a depletion of antibodies capable of blocking ligand binding, adsorption by the everted membrane preparation led to a relative enrichment of blocking antibodies. These results are interpreted as indicative of distinct antigenic determinants of the receptor being present on the two faces of the membrane and demonstrate a transbilayer disposition of the asialoglycoprotein receptor.

Full text

PDF
1557

Selected References

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

  1. Anderson R. G., Brown M. S., Goldstein J. L. Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblasts. Cell. 1977 Mar;10(3):351–364. doi: 10.1016/0092-8674(77)90022-8. [DOI] [PubMed] [Google Scholar]
  2. Anderson R. G., Goldstein J. L., Brown M. S. A mutation that impairs the ability of lipoprotein receptors to localise in coated pits on the cell surface of human fibroblasts. Nature. 1977 Dec 22;270(5639):695–699. doi: 10.1038/270695a0. [DOI] [PubMed] [Google Scholar]
  3. Ashwell G., Morell A. G. The role of surface carbohydrates in the hepatic recognition and transport of circulating glycoproteins. Adv Enzymol Relat Areas Mol Biol. 1974;41(0):99–128. doi: 10.1002/9780470122860.ch3. [DOI] [PubMed] [Google Scholar]
  4. Brown M. S., Goldstein J. L. Analysis of a mutant strain of human fibroblasts with a defect in the internalization of receptor-bound low density lipoprotein. Cell. 1976 Dec;9(4 Pt 2):663–674. doi: 10.1016/0092-8674(76)90130-6. [DOI] [PubMed] [Google Scholar]
  5. Chan S. H., Schatz G. Use of antibodies for studying the sidedness of membrane components. Methods Enzymol. 1979;56:223–228. doi: 10.1016/0076-6879(79)56024-8. [DOI] [PubMed] [Google Scholar]
  6. Cohen C. M., Kalish D. I., Jacobson B. S., Branton D. Membrane isolation on polylysine-coated beads. Plasma membrane from HeLa cells. J Cell Biol. 1977 Oct;75(1):119–134. doi: 10.1083/jcb.75.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cohen C. M., Kramer R. M., Branton D. Transbilayer mapping of membrane proteins using membranes isolated on polylysine-coated polyacrylamide beads. Biochim Biophys Acta. 1980 Mar 27;597(1):29–40. doi: 10.1016/0005-2736(80)90147-9. [DOI] [PubMed] [Google Scholar]
  8. Cuatrecasas P. Membrane receptors. Annu Rev Biochem. 1974;43(0):169–214. doi: 10.1146/annurev.bi.43.070174.001125. [DOI] [PubMed] [Google Scholar]
  9. Goldstein J. L., Anderson R. G., Brown M. S. Coated pits, coated vesicles, and receptor-mediated endocytosis. Nature. 1979 Jun 21;279(5715):679–685. doi: 10.1038/279679a0. [DOI] [PubMed] [Google Scholar]
  10. Gregoriadis G., Morell A. G., Sternlieb I., Scheinberg I. H. Catabolism of desialylated ceruloplasmin in the liver. J Biol Chem. 1970 Nov 10;245(21):5833–5837. [PubMed] [Google Scholar]
  11. Guidotti G. Membrane proteins. Annu Rev Biochem. 1972;41:731–752. doi: 10.1146/annurev.bi.41.070172.003503. [DOI] [PubMed] [Google Scholar]
  12. HELMKAMP R. W., GOODLAND R. L., BALE W. F., SPAR I. L., MUTSCHLER L. E. High specific activity iodination of gamma-globulin with iodine-131 monochloride. Cancer Res. 1960 Nov;20:1495–1500. [PubMed] [Google Scholar]
  13. Jacobson B. S., Branton D. Plasma membrane: rapid isolation and exposure of the cytoplasmic surface by use of positively charged beads. Science. 1977 Jan 21;195(4275):302–304. doi: 10.1126/science.831278. [DOI] [PubMed] [Google Scholar]
  14. Kalish D. I., Cohen C. M., Jacobson B. S., Branton D. Membrane isolation on polylysine-coated glass beads. Asymmetry of bound membrane. Biochim Biophys Acta. 1978 Jan 4;506(1):97–110. doi: 10.1016/0005-2736(78)90437-6. [DOI] [PubMed] [Google Scholar]
  15. Kawasaki T., Ashwell G. Isolation and characterization of an avian hepatic binding protein specific for N-acetylglucosamine-terminated glycoproteins. J Biol Chem. 1977 Sep 25;252(18):6536–6543. [PubMed] [Google Scholar]
  16. 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]
  17. Marchesi V. T., Furthmayr H., Tomita M. The red cell membrane. Annu Rev Biochem. 1976;45:667–698. doi: 10.1146/annurev.bi.45.070176.003315. [DOI] [PubMed] [Google Scholar]
  18. Morell A. G., Gregoriadis G., Scheinberg I. H., Hickman J., Ashwell G. The role of sialic acid in determining the survival of glycoproteins in the circulation. J Biol Chem. 1971 Mar 10;246(5):1461–1467. [PubMed] [Google Scholar]
  19. Nicolson G. L. Transmembrane control of the receptors on normal and tumor cells. I. Cytoplasmic influence over surface components. Biochim Biophys Acta. 1976 Apr 13;457(1):57–108. doi: 10.1016/0304-4157(76)90014-9. [DOI] [PubMed] [Google Scholar]
  20. Pricer W. E., Jr, Ashwell G. Subcellular distribution of a mammalian hepatic binding protein specific for asialoglycoproteins. J Biol Chem. 1976 Dec 10;251(23):7539–7544. [PubMed] [Google Scholar]
  21. Pricer W. E., Jr, Ashwell G. The binding of desialylated glycoproteins by plasma membranes of rat liver. J Biol Chem. 1971 Aug 10;246(15):4825–4833. [PubMed] [Google Scholar]
  22. Regoeczi E., Taylor P., Hatton M. W., Wong K. L., Koj A. Distinction between binding and endocytosis of human asialo-transferrin by the rat liver. Biochem J. 1978 Jul 15;174(1):171–178. doi: 10.1042/bj1740171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rothman J. E., Lenard J. Membrane asymmetry. Science. 1977 Feb 25;195(4280):743–753. doi: 10.1126/science.402030. [DOI] [PubMed] [Google Scholar]
  24. Singer S. J. Molecular biology of cellular membranes with applications to immunology. Adv Immunol. 1974;19(0):1–66. doi: 10.1016/s0065-2776(08)60251-5. [DOI] [PubMed] [Google Scholar]
  25. Steer C. J., Ashwell G. Studies on a mammalian hepatic binding protein specific for asialoglycoproteins. Evidence for receptor recycling in isolated rat hepatocytes. J Biol Chem. 1980 Apr 10;255(7):3008–3013. [PubMed] [Google Scholar]
  26. Stockert R. J., Howard D. J., Morell A. G., Scheinberg I. H. Functional segregation of hepatic receptors for asialoglycoproteins during endocytosis. J Biol Chem. 1980 Oct 10;255(19):9028–9029. [PubMed] [Google Scholar]
  27. Tanabe T., Pricer W. E., Jr, Ashwell G. Subcellular membrane topology and turnover of a rat hepatic binding protein specific for asialoglycoproteins. J Biol Chem. 1979 Feb 25;254(4):1038–1043. [PubMed] [Google Scholar]
  28. Tolleshaug H., Berg T., Nilsson M., Norum K. R. Uptake and degradation of 125I-labelled asialo-fetuin by isolated rat hepatocytes. Biochim Biophys Acta. 1977 Aug 25;499(1):73–84. doi: 10.1016/0304-4165(77)90230-6. [DOI] [PubMed] [Google Scholar]
  29. Wall D. A., Wilson G., Hubbard A. L. The galactose-specific recognition system of mammalian liver: the route of ligand internalization in rat hepatocytes. Cell. 1980 Aug;21(1):79–93. doi: 10.1016/0092-8674(80)90116-6. [DOI] [PubMed] [Google Scholar]
  30. Weigel P. H. Characterization of the asialoglycoprotein receptor on isolated rat hepatocytes. J Biol Chem. 1980 Jul 10;255(13):6111–6120. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES