Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1995 Feb 15;306(Pt 1):129–134. doi: 10.1042/bj3060129

Identification of a novel form of the alpha 3 integrin subunit: covalent association with transferrin receptor.

M Coppolino 1, M Migliorini 1, W S Argraves 1, S Dedhar 1
PMCID: PMC1136491  PMID: 7864799

Abstract

The alpha 3 beta 1 integrin is a cell-surface receptor for laminin, entactin, collagen, fibronectin and epiligrin. On some prostatic-carcinoma cell lines that express the alpha 3 beta 1 heterodimer we have identified a novel form of the alpha 3 subunit. Whereas the prototypic alpha 3 subunit has a molecular mass of approximately 155 kDa, we have isolated a approximately 225 kDa protein (p225) which is recognized by monoclonal antibodies to the alpha 3 subunit. Protein sequence analysis revealed that p225 consists of two polypeptides, namely integrin alpha 3 heavy chain (approximately 130 kDa) disulphide-bonded to a monomer of the transferrin receptor (approximately 95 kDa) instead of the typical alpha 3 light chain (approximately 25 kDa). The p225 seems to be directly associated with beta 1 subunit, since it was immunoprecipitable with anti-(beta 1 subunit) antibodies. The association of transferrin receptor and integrin alpha 3 was apparently not the result of spurious disulphide-bond formation occurring during the protein purification, as iodoacetamide and GSH did not block the formation of the complex. The transferrin receptor is normally a homodimer that is involved in the internalization of iron-bound transferrin into cells and can be expressed at relatively high levels in the cell lines which we have studied. The p225 is not found on all cell types examined to date and therefore it may represent a unique complex between the integrin alpha 3 subunit and the transferrin receptor, a covalent association which may play a role in the adherence and/or proliferation of some types of tumour cells.

Full text

PDF
129

Images in this article

131Figure 1
on p.131
131Figure 3
on p.131
132Figure 4
on p.132
132Figure 5
on p.132

Selected References

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

  1. Aebersold R. H., Leavitt J., Saavedra R. A., Hood L. E., Kent S. B. Internal amino acid sequence analysis of proteins separated by one- or two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose. Proc Natl Acad Sci U S A. 1987 Oct;84(20):6970–6974. doi: 10.1073/pnas.84.20.6970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albelda S. M., Buck C. A. Integrins and other cell adhesion molecules. FASEB J. 1990 Aug;4(11):2868–2880. [PubMed] [Google Scholar]
  3. Carter W. G., Ryan M. C., Gahr P. J. Epiligrin, a new cell adhesion ligand for integrin alpha 3 beta 1 in epithelial basement membranes. Cell. 1991 May 17;65(4):599–610. doi: 10.1016/0092-8674(91)90092-d. [DOI] [PubMed] [Google Scholar]
  4. Collawn J. F., Lai A., Domingo D., Fitch M., Hatton S., Trowbridge I. S. YTRF is the conserved internalization signal of the transferrin receptor, and a second YTRF signal at position 31-34 enhances endocytosis. J Biol Chem. 1993 Oct 15;268(29):21686–21692. [PubMed] [Google Scholar]
  5. Cooper H. M., Tamura R. N., Quaranta V. The major laminin receptor of mouse embryonic stem cells is a novel isoform of the alpha 6 beta 1 integrin. J Cell Biol. 1991 Nov;115(3):843–850. doi: 10.1083/jcb.115.3.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dautry-Varsat A., Ciechanover A., Lodish H. F. pH and the recycling of transferrin during receptor-mediated endocytosis. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2258–2262. doi: 10.1073/pnas.80.8.2258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dedhar S., Gray V., Robertson K., Saulnier R. Identification and characterization of a novel high-molecular-weight form of the integrin alpha 3 subunit. Exp Cell Res. 1992 Nov;203(1):270–275. doi: 10.1016/0014-4827(92)90064-f. [DOI] [PubMed] [Google Scholar]
  8. Dedhar S., Haqq C., Gray V. Specific overproduction of very late antigen 1 integrin in two human neuroblastoma cell lines selected for resistance to detachment by an Arg-Gly-Asp-containing synthetic peptide. J Biol Chem. 1989 Mar 25;264(9):4832–4836. [PubMed] [Google Scholar]
  9. Dedhar S. Integrins and tumor invasion. Bioessays. 1990 Dec;12(12):583–590. doi: 10.1002/bies.950121205. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Hemler M. E. VLA proteins in the integrin family: structures, functions, and their role on leukocytes. Annu Rev Immunol. 1990;8:365–400. doi: 10.1146/annurev.iy.08.040190.002053. [DOI] [PubMed] [Google Scholar]
  12. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  13. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  14. JANDL J. H., KATZ J. H. The plasma-to-cell cycle of transferrin. J Clin Invest. 1963 Mar;42:314–326. doi: 10.1172/JCI104718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kantor R. R., Mattes M. J., Lloyd K. O., Old L. J., Albino A. P. Biochemical analysis of two cell surface glycoprotein complexes, very common antigen 1 and very common antigen 2. Relationship to very late activation T cell antigens. J Biol Chem. 1987 Nov 5;262(31):15158–15165. [PubMed] [Google Scholar]
  16. Keer H. N., Kozlowski J. M., Tsai Y. C., Lee C., McEwan R. N., Grayhack J. T. Elevated transferrin receptor content in human prostate cancer cell lines assessed in vitro and in vivo. J Urol. 1990 Feb;143(2):381–385. doi: 10.1016/s0022-5347(17)39970-6. [DOI] [PubMed] [Google Scholar]
  17. Larrick J. W., Cresswell P. Modulation of cell surface iron transferrin receptors by cellular density and state of activation. J Supramol Struct. 1979;11(4):579–586. doi: 10.1002/jss.400110415. [DOI] [PubMed] [Google Scholar]
  18. Lebien T. W., Boué D. R., Bradley J. G., Kersey J. H. Antibody affinity may influence antigenic modulation of the common acute lymphoblastic leukemia antigen in vitro. J Immunol. 1982 Nov;129(5):2287–2292. [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. Musgrove E., Rugg C., Taylor I., Hedley D. Transferrin receptor expression during exponential and plateau phase growth of human tumour cells in culture. J Cell Physiol. 1984 Jan;118(1):6–12. doi: 10.1002/jcp.1041180103. [DOI] [PubMed] [Google Scholar]
  21. Neckers L. M., Trepel J. B. Transferrin receptor expression and the control of cell growth. Cancer Invest. 1986;4(5):461–470. doi: 10.3109/07357908609017524. [DOI] [PubMed] [Google Scholar]
  22. Ruoslahti E. Integrins. J Clin Invest. 1991 Jan;87(1):1–5. doi: 10.1172/JCI114957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Schwartz M. A. Signaling by integrins: implications for tumorigenesis. Cancer Res. 1993 Apr 1;53(7):1503–1506. [PubMed] [Google Scholar]
  25. Sonnenberg A., Modderman P. W., Hogervorst F. Laminin receptor on platelets is the integrin VLA-6. Nature. 1988 Dec 1;336(6198):487–489. doi: 10.1038/336487a0. [DOI] [PubMed] [Google Scholar]
  26. Sriramarao P., Steffner P., Gehlsen K. R. Biochemical evidence for a homophilic interaction of the alpha 3 beta 1 integrin. J Biol Chem. 1993 Oct 15;268(29):22036–22041. [PubMed] [Google Scholar]
  27. Symington B. E., Takada Y., Carter W. G. Interaction of integrins alpha 3 beta 1 and alpha 2 beta 1: potential role in keratinocyte intercellular adhesion. J Cell Biol. 1993 Jan;120(2):523–535. doi: 10.1083/jcb.120.2.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Takada Y., Murphy E., Pil P., Chen C., Ginsberg M. H., Hemler M. E. Molecular cloning and expression of the cDNA for alpha 3 subunit of human alpha 3 beta 1 (VLA-3), an integrin receptor for fibronectin, laminin, and collagen. J Cell Biol. 1991 Oct;115(1):257–266. doi: 10.1083/jcb.115.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tamura R. N., Cooper H. M., Collo G., Quaranta V. Cell type-specific integrin variants with alternative alpha chain cytoplasmic domains. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10183–10187. doi: 10.1073/pnas.88.22.10183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tamura R. N., Rozzo C., Starr L., Chambers J., Reichardt L. F., Cooper H. M., Quaranta V. Epithelial integrin alpha 6 beta 4: complete primary structure of alpha 6 and variant forms of beta 4. J Cell Biol. 1990 Oct;111(4):1593–1604. doi: 10.1083/jcb.111.4.1593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Trowbridge I. S., Lopez F. Monoclonal antibody to transferrin receptor blocks transferrin binding and inhibits human tumor cell growth in vitro. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1175–1179. doi: 10.1073/pnas.79.4.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. van Kuppevelt T. H., Languino L. R., Gailit J. O., Suzuki S., Ruoslahti E. An alternative cytoplasmic domain of the integrin beta 3 subunit. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5415–5418. doi: 10.1073/pnas.86.14.5415. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES