Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

The intrinsic factor–vitamin B12 receptor, cubilin, is a high-affinity apolipoprotein A-I receptor facilitating endocytosis of high-density lipoprotein

Nature Medicine volume 5, pages 656–661 (1999)Cite this article

Abstract

Cubilin is the intestinal receptor for the endocytosis of intrinsic factor–vitamin B12. However, several lines of evidence, including a high expression in kidney and yolk sac, indicate it may have additional functions. We isolated apolipoprotein A-I (apoA-I), the main protein of high-density lipoprotein (HDL), using cubilin affinity chromatography. Surface plasmon resonance analysis demonstrated a high-affinity binding of apoA-I and HDL to cubilin, and cubilin-expressing yolk sac cells showed efficient 125I-HDL endocytosis that could be inhibited by IgG antibodies against apoA-I and cubilin. The physiological relevance of the cubilin–apoA-I interaction was further emphasized by urinary apoA-I loss in some known cases of functional cubilin deficiency. Therefore, cubilin is a receptor in epithelial apoA-I/HDL metabolism.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Identification of apoA-I as a cubilin ligand by affinity chromatography.
Figure 2: Binding of apoA-I and apoA-I-HDL to cubilin.
Figure 3: Uptake and degradation of 125I-HDL and 125I-LDL in cultured rat yolk sac epithelial cells.
Figure 4: Saturation curves for cell association of 125I-HDL in rat yolk sac epithelial cells at 4 °C (a, 4 h) and 37 °C (b, 30 min).
Figure 5: Cell association and degradation of 125I-HDL in cultured rat yolk sac cells.
Figure 6: Renal handling of apoA-I and urinary excretion of apoA-I in patients and dogs with Imerslund-Gräsbeck (I.G.) syndrome.

Similar content being viewed by others

References

  1. Birn, H. et al. Characterization of an epithelial approximately 460-kDa protein that facilitates endocytosis of intrinsic factor-vitamin B12 and binds receptor-associated protein. J. Biol. Chem. 272, 26497– 26504 (1997).

    Article  CAS  Google Scholar 

  2. Moestrup, S.K. et al. The intrinsic factor-vitamin B12 receptor and target of teratogenic antibodies is a megalin-binding peripheral membrane protein with homology to developmental proteins. J. Biol. Chem. 273, 5235–5242 (1998).

    Article  CAS  Google Scholar 

  3. Kozyraki, R. et al. The human intrinsic factor-vitamin B12 receptor, cubilin: molecular characterization and chromosomal mapping of the gene to 10p within the autosomal recessive megaloblastic anemia (MGA1) region. Blood 91, 3593–3600 ( 1998).

    CAS  PubMed  Google Scholar 

  4. Aminoff, M. et al. Mutations in the CUBN gene encoding the intrinsic factor vitamin B12 receptor, cubilin cause hereditary megaloblastic anemia 1 (MGA-1) Nature Genet. 21, 309–313 (1999).

    Article  CAS  Google Scholar 

  5. Fyfe, J.C., Ramanujam, K.S., Ramaswamy, K., Patterson, D.F., & Seetharam, B. Defective brush-border expression of intrinsic factor-cobalamin receptor in canine inherited intestinal malabsorption. J. Biol. Chem. 266, 4489– 94 (1991).

    CAS  PubMed  Google Scholar 

  6. Sahali, D. et al. Characterization of a 280-kD protein restricted to the coated pits of the renal brush border and the epithelial cells of the yolk sac. Teratogenic effect of the specific monoclonal antibodies. J. Exp. Med. 167, 213–218 ( 1988).

    Article  CAS  Google Scholar 

  7. Seetharam, B., Christensen, E.I., Moestrup, S.K., Hammond, T.G. & Verroust, P.J. Identification of rat yolk sac target protein of teratogenic antibodies, gp280, as intrinsic factor-cobalamin receptor. J. Clin. Invest. 99, 2317– 2322 (1997).

    Article  CAS  Google Scholar 

  8. Imerslund O. Idiopathic chronic megaloblastic anemia in children. Acta Paed. Scand. 49, 1–115 ( 1960).

    Google Scholar 

  9. Gräsbeck R, Gordin R, Kantero I & Kuhlback B : Selective vitamin B12 malabsorption and proteinuria in young people. Acta Med. Scand. 167, 289–296 (1960).

    Article  Google Scholar 

  10. Brown, M.S. & Goldstein, J.L. A receptor-mediated pathway for cholesterol homeostasis. Science 232, 34–47 (1986).

    Article  CAS  Google Scholar 

  11. Acton, S. et al.. Identification of scavenger receptor SR-BI as a high density lipoprotein receptor. Science 271, 518– 520 (1996).

    Article  CAS  Google Scholar 

  12. Le Panse, .S. et al. Immunofunctional properties of a yolk sac epithelial cell line expressing two proteins gp280 and gp330 of the intermicrovillar area of proximal tubule cells: inhibition of endocytosis by the specific antibodies. Eur. J. Cell Biol. 67, 120– 129 (1995).

    CAS  PubMed  Google Scholar 

  13. Wyne, K.L. & Woollett, L.A. Transport of maternal LDL and HDL to the fetal membranes and placenta of the Golden Syrian hamster is mediated by receptor-dependent and receptor-independent processes. J. Lipid. Res. 39, 518–530 ( 1998).

    CAS  PubMed  Google Scholar 

  14. Moestrup, S.K. et al. β2-Glycoprotein-I (apolipoprotein-H) and β 2-Glycoprotein-I-phospholipid complex harbor a recognition site for the endocytic receptor megalin. J. Clin. Invest. 102 , 902–909 (1998).

    Article  CAS  Google Scholar 

  15. Stefansson, S., Chappell, D.A., Argraves, K.M., Strickland, D.K. & Argraves, W.S. Glycoprotein 330/low density lipoprotein receptor-related protein-2 mediates endocytosis of low density lipoproteins via interaction with apolipoprotein B100. J. Biol. Chem. 279, 19417–19421(1995).

    Article  Google Scholar 

  16. Glass, C., Pittman, R.C., Civen, M. & Steinberg, D. Uptake of high-density lipoprotein-associated apolipoprotein A-I and cholesterol esters by 16 tissues of the rat in vivo and by adrenal cells and hepatocytes in vitro. J. Biol. Chem. 260, 744–750 (1985).

    CAS  PubMed  Google Scholar 

  17. Willnow, T.E., Goldstein, J.L., Orth, K., Brown, M.S. & Herz, J. Low density lipoprotein receptor-related protein and gp330 bind similar ligands, including plasminogen activator-inhibitor complexes and lactoferrin, an inhibitor of chylomicron remnant clearance. J. Biol. Chem. 267, 26172– 26180 (1992).

    CAS  PubMed  Google Scholar 

  18. Kounnas, M.Z. et al. Identification of glycoprotein 330 as an endocytic receptor for apolipoprotein J/clusterin. J. Biol. Chem. 270, 13070–13075 (1995).

    Article  CAS  Google Scholar 

  19. Kozarksky, K.F. et al. Overexpression of the HDL receptor SR-BI alters plasma HDL and bile cholesterol levels. Nature 387, 414–417 (1997).

    Article  Google Scholar 

  20. Saku K., Reddy, G.S., Hynd, B.A. & Kashyap. M. Renal handling of high-density lipoprotein by isolated perfused kidneys. Metabolism 33, 432–438 ( 1984).

    Article  CAS  Google Scholar 

  21. Farese, R.V., Jr. & Herz, J. Cholesterol metabolism and embryogenesis. Trends Genet. 14, 115–120 (1998).

    Article  CAS  Google Scholar 

  22. Kolf-Clauw, M. et al. Inhibition of 7-dehydrocholesterol reductase by the teratogen AY9944: a rat model for Smith-Lemli-Opitz syndrome. Teratology 54, 115–125 ( 1996).

    Article  CAS  Google Scholar 

  23. Cooper, M.K., Porter, J.A, Young. K.E., Beachy, P.A. Teratogen-mediated inhibition of target tissue response to Shh signaling. Science 280 , 1603–1607. (1998).

    Article  CAS  Google Scholar 

  24. Farese, R.V. Jr. et al. A novel function for apolipoprotein B: lipoprotein synthesis in the yolk sac is critical for maternal-fetal lipid transport in mice. J. Lipid Res. 37, 347–360 (1996).

    CAS  PubMed  Google Scholar 

  25. Farese, R.V. Jr., Ruland, S.L., Flynn, L.M., Stokowski, R.P. & Young, S.G. Knockout of the mouse apolipoprotein B gene results in embryonic lethality in homozygotes and protection against diet-induced hypercholesterolemia in heterozygotes. Proc. Natl. Acad. Sci. USA 92, 1774–1778 (1995).

    Article  CAS  Google Scholar 

  26. Homanics, G.E. et al. Exencephaly and hydrocephaly in mice with targeted modification of the apolipoprotein B (Apob) gene. Teratology 51, 1–10 (1995).

    Article  CAS  Google Scholar 

  27. Willnow, T.E. et al. Defective forebrain development in mice lacking gp330/megalin. Proc. Natl. Acad. Sci. USA 93, 8460– 8464 (1996).

    Article  CAS  Google Scholar 

  28. Sahali, D. et al. Comparative immunochemistry and ontogeny of two closely related coated pit proteins. The 280-kd target of teratogenic antibodies and the 330-kd target of nephritogenic antibodies. Am. J. Pathol. 142, 1654–1667 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Plump, A.S. et al. ApoA-I knockout mice:characterization of HDL metabolism in homozygotes and identification of a post-RNA mechanism of apoA-I upregulation in heterozygotes. J. Lipid. Res. 36, 1933 –1047 (1997).

    Google Scholar 

  30. Rigotti, A. et al. Scavenger receptor BI—a cell surface receptor for high density lipoprotein. Curr. Opin. Lipidol. 8, 181–188 (1997).

    Article  CAS  Google Scholar 

  31. Hatzopoulos, A.K., Rigotti, A., Rosenberg, R.D & Krieger, M. Temporal and spatial pattern of expression of the HDL receptor SR-BI during murine embryogenesis. J. Lipid Res. 39, 495–508 (1998).

    CAS  PubMed  Google Scholar 

  32. Hatch, F.T. & Lees, R.S. . Practical methods for plasma lipoprotein analysis. Adv. Lipid. Res. 6, 1– 68 (1968).

    Article  CAS  Google Scholar 

  33. Moestrup, S.K. et al.. Megalin-mediated endocytosis of transcobalamin-vitamin-B12 complexes suggests a role of the receptor in vitamin-B12 homeostasis. Proc. Natl. Acad. Sci. USA 93, 8612– 8617 (1996).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The technical assistance of K. Lassen is acknowledged. The patients are thanked for their cooperation and S. Lindh, for sample collection. U. Gerdes and J. Møller are thanked for the cholesterol and creatinin measurements. J. Gliemann is thanked for comments on the manuscript. The work was financially supported by the Novo Nordisk Foundation, the Danish Medical Research Council, the Danish Biomembrane Center, Aarhus University Research Foundation, the Velux Foundation, a contract from ARC (9914, le Ministére des Affaires Etrangéres, France, NIH grant DK45341 (J.C.F.) and National Cancer Institute grant P30 CA16058.

Author information

Authors and Affiliations

  1. Department of Medical Biochemistry, University of Aarhus, Denmark

    Renata Kozyraki, Mette Kristiansen, Christian Jacobsen & Søren K. Moestrup

  2. Department of Microbiology, Michigan State University, East Lansing, Michigan, USA

    John Fyfe

  3. Department of Medicine and Cardiology A, Aarhus University Hospital, Denmark

    Christian Gerdes

  4. Department of Cell Biology, Institute of Anatomy, University of Aarhus, Denmark

    Shiying Cui & Erik Ilsø Christensen

  5. Division of Human Cancer Genetics, Department of Microbiology and Immunology, Comprehensive Cancer Center, Ohio state University, Columbus, Ohio, USA

    Maria Aminoff, Albert de la Chapelle & Ralf Krahe

  6. Folkhälsan Institute of Genetics, University of Helsinki, Finland

    Maria Aminoff, Albert de la Chapelle & Ralf Krahe

  7. Institut National de la Santé et de la Recherche Medicale, U489, Hopital Tenon, Paris, France

    Pierre J. Verroust

Authors
  1. Renata Kozyraki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John Fyfe
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mette Kristiansen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christian Gerdes
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Christian Jacobsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shiying Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Erik Ilsø Christensen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Maria Aminoff
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Albert de la Chapelle
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Ralf Krahe
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Pierre J. Verroust
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Søren K. Moestrup
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Søren K. Moestrup.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kozyraki, R., Fyfe, J., Kristiansen, M. et al. The intrinsic factor–vitamin B12 receptor, cubilin, is a high-affinity apolipoprotein A-I receptor facilitating endocytosis of high-density lipoprotein. Nat Med 5, 656–661 (1999). https://doi.org/10.1038/9504

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/9504

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing