Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1989 Jun 1;108(6):2127–2135. doi: 10.1083/jcb.108.6.2127

Concentration of transferrin receptor in human placental coated vesicles

PMCID: PMC2115582  PMID: 2567737

Abstract

Coated vesicles were purified from human placenta by sedimentation, isopycnic centrifugation, and gel filtration. Quantitative Western blotting of the endogenous transferrin receptor (tfR) demonstrated the presence, on average, of roughly one receptor per vesicle. TfR appeared undersaturated with transferrin. After solubilizing vesicles in nonionic detergent, we looked for evidence of tfR interactions with other proteins. Solubilized tfR had an unexpectedly high mobility by gel filtration, apparently resulting from its self-association. This property was not seen in purified tfR or in tfR from a different cell fraction. The tfR complexes, though noncovalent, were largely resistant to conditions that disassemble coat proteins, and they did not appear to contain any other protein species.

Full Text

The Full Text of this article is available as a PDF (2.8 MB).

Selected References

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

  1. Altstiel L., Branton D. Fusion of coated vesicles with lysosomes: measurement with a fluorescence assay. Cell. 1983 Mar;32(3):921–929. doi: 10.1016/0092-8674(83)90077-6. [DOI] [PubMed] [Google Scholar]
  2. Anderson G. J., Mackerras A., Powell L. W., Halliday J. W. Improved purification of the human placental transferrin receptor and a novel immunoradiometric assay for receptor protein. Biochim Biophys Acta. 1986 Nov 19;884(2):225–233. doi: 10.1016/0304-4165(86)90167-4. [DOI] [PubMed] [Google Scholar]
  3. Booth A. G., Wilson M. J. Human placental coated vesicles contain receptor-bound transferrin. Biochem J. 1981 Apr 15;196(1):355–362. doi: 10.1042/bj1960355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bretscher M. S., Thomson J. N., Pearse B. M. Coated pits act as molecular filters. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4156–4159. doi: 10.1073/pnas.77.7.4156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ciechanover A., Schwartz A. L., Lodish H. F. The asialoglycoprotein receptor internalizes and recycles independently of the transferrin and insulin receptors. Cell. 1983 Jan;32(1):267–275. doi: 10.1016/0092-8674(83)90517-2. [DOI] [PubMed] [Google Scholar]
  6. Crowther R. A., Finch J. T., Pearse B. M. On the structure of coated vesicles. J Mol Biol. 1976 Jun 5;103(4):785–798. doi: 10.1016/0022-2836(76)90209-6. [DOI] [PubMed] [Google Scholar]
  7. Crowther R. A., Pearse B. M. Assembly and packing of clathrin into coats. J Cell Biol. 1981 Dec;91(3 Pt 1):790–797. doi: 10.1083/jcb.91.3.790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davis C. G., Lehrman M. A., Russell D. W., Anderson R. G., Brown M. S., Goldstein J. L. The J.D. mutation in familial hypercholesterolemia: amino acid substitution in cytoplasmic domain impedes internalization of LDL receptors. Cell. 1986 Apr 11;45(1):15–24. doi: 10.1016/0092-8674(86)90533-7. [DOI] [PubMed] [Google Scholar]
  9. Enns C. A., Sussman H. H. Physical characterization of the transferrin receptor in human placentae. J Biol Chem. 1981 Oct 10;256(19):9820–9823. [PubMed] [Google Scholar]
  10. Forgac M., Cantley L., Wiedenmann B., Altstiel L., Branton D. Clathrin-coated vesicles contain an ATP-dependent proton pump. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1300–1303. doi: 10.1073/pnas.80.5.1300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grasberger B., Minton A. P., DeLisi C., Metzger H. Interaction between proteins localized in membranes. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6258–6262. doi: 10.1073/pnas.83.17.6258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Heuser J. Three-dimensional visualization of coated vesicle formation in fibroblasts. J Cell Biol. 1980 Mar;84(3):560–583. doi: 10.1083/jcb.84.3.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hope I. A., Struhl K. Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of yeast. Cell. 1986 Sep 12;46(6):885–894. doi: 10.1016/0092-8674(86)90070-x. [DOI] [PubMed] [Google Scholar]
  14. Hopkins C. R. Intracellular routing of transferrin and transferrin receptors in epidermoid carcinoma A431 cells. Cell. 1983 Nov;35(1):321–330. doi: 10.1016/0092-8674(83)90235-0. [DOI] [PubMed] [Google Scholar]
  15. Iacopetta B. J., Rothenberger S., Kühn L. C. A role for the cytoplasmic domain in transferrin receptor sorting and coated pit formation during endocytosis. Cell. 1988 Aug 12;54(4):485–489. doi: 10.1016/0092-8674(88)90069-4. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lewis R. V., Roberts M. F., Dennis E. A., Allison W. S. Photoactivated heterobifunctional cross-linking reagents which demonstrate the aggregation state of phospholipase A2. Biochemistry. 1977 Dec 13;16(25):5650–5654. doi: 10.1021/bi00644a041. [DOI] [PubMed] [Google Scholar]
  18. Ma J., Ptashne M. A new class of yeast transcriptional activators. Cell. 1987 Oct 9;51(1):113–119. doi: 10.1016/0092-8674(87)90015-8. [DOI] [PubMed] [Google Scholar]
  19. McClelland A., Kühn L. C., Ruddle F. H. The human transferrin receptor gene: genomic organization, and the complete primary structure of the receptor deduced from a cDNA sequence. Cell. 1984 Dec;39(2 Pt 1):267–274. doi: 10.1016/0092-8674(84)90004-7. [DOI] [PubMed] [Google Scholar]
  20. Moore M. S., Mahaffey D. T., Brodsky F. M., Anderson R. G. Assembly of clathrin-coated pits onto purified plasma membranes. Science. 1987 May 1;236(4801):558–563. doi: 10.1126/science.2883727. [DOI] [PubMed] [Google Scholar]
  21. Pearse B. M. Coated vesicles from human placenta carry ferritin, transferrin, and immunoglobulin G. Proc Natl Acad Sci U S A. 1982 Jan;79(2):451–455. doi: 10.1073/pnas.79.2.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pearse B. M., Crowther R. A. Structure and assembly of coated vesicles. Annu Rev Biophys Biophys Chem. 1987;16:49–68. doi: 10.1146/annurev.bb.16.060187.000405. [DOI] [PubMed] [Google Scholar]
  23. Pearse B. M. Receptors compete for adaptors found in plasma membrane coated pits. EMBO J. 1988 Nov;7(11):3331–3336. doi: 10.1002/j.1460-2075.1988.tb03204.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schneider C., Newman R. A., Sutherland D. R., Asser U., Greaves M. F. A one-step purification of membrane proteins using a high efficiency immunomatrix. J Biol Chem. 1982 Sep 25;257(18):10766–10769. [PubMed] [Google Scholar]
  25. Schneider C., Owen M. J., Banville D., Williams J. G. Primary structure of human transferrin receptor deduced from the mRNA sequence. Nature. 1984 Oct 18;311(5987):675–678. doi: 10.1038/311675b0. [DOI] [PubMed] [Google Scholar]
  26. Schneider C., Sutherland R., Newman R., Greaves M. Structural features of the cell surface receptor for transferrin that is recognized by the monoclonal antibody OKT9. J Biol Chem. 1982 Jul 25;257(14):8516–8522. [PubMed] [Google Scholar]
  27. Seligman P. A., Schleicher R. B., Allen R. H. Isolation and characterization of the transferrin receptor from human placenta. J Biol Chem. 1979 Oct 25;254(20):9943–9946. [PubMed] [Google Scholar]
  28. Steven A. C., Hainfeld J. F., Wall J. S., Steer C. J. Mass distributions of coated vesicles isolated from liver and brain: analysis by scanning transmission electron microscopy. J Cell Biol. 1983 Dec;97(6):1714–1723. doi: 10.1083/jcb.97.6.1714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Turkewitz A. P., Amatruda J. F., Borhani D., Harrison S. C., Schwartz A. L. A high yield purification of the human transferrin receptor and properties of its major extracellular fragment. J Biol Chem. 1988 Jun 15;263(17):8318–8325. [PubMed] [Google Scholar]
  30. Turkewitz A. P., Schwartz A. L., Harrison S. C. A pH-dependent reversible conformational transition of the human transferrin receptor leads to self-association. J Biol Chem. 1988 Nov 5;263(31):16309–16315. [PubMed] [Google Scholar]
  31. Watts C. In situ 125I-labelling of endosome proteins with lactoperoxidase conjugates. EMBO J. 1984 Sep;3(9):1965–1970. doi: 10.1002/j.1460-2075.1984.tb02077.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yamashiro D. J., Maxfield F. R. Kinetics of endosome acidification in mutant and wild-type Chinese hamster ovary cells. J Cell Biol. 1987 Dec;105(6 Pt 1):2713–2721. doi: 10.1083/jcb.105.6.2713. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES