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. 1987 Jul 1;105(1):589–598. doi: 10.1083/jcb.105.1.589

Two distinct cell-binding domains in laminin can independently promote nonneuronal cell adhesion and spreading [published erratum appears in J Cell Biol 1989 Jun;108(6):following 2546]

PMCID: PMC2114908  PMID: 3038930

Abstract

Two subfragments of laminin, E8, a major part of the long arm, and E1- 4, the three short arms, promote cell adhesion and spreading. Three distinct types of adhesive behavior are seen in short term (1 h) assays, typified by secondary murine fibroblasts, adherent only on fibronectin; secondary murine myoblasts, adherent on fibronectin, laminin, and the E8 fragment; and Rugli human glioblastoma cells, adherent on fibronectin, laminin, E8, and E1-4. E8-specific polyclonal antibodies block myoblast adhesion to E8 and to laminin with identical concentration dependence; Rugli binding to E8 but not to laminin is also totally blocked by these antibodies. Heating of E8 and laminin to approximately 60 degrees C abolishes cell attachment-promoting activity for myoblasts. Adhesion of Rugli cells to E8 is also lost, but on laminin the attachment-promoting activity remains constant. This is due to an increase in the activity of E1-4 fragment as it is heated. Thus, major sites for initial cell adhesion to and spreading on laminin lie within the E8 and E1-4 fragments, but not all cells binding to laminin will bind to both fragments. These data may tentatively be explained by the existence of more than one type of receptor for laminin at the cell surface; one is needed for each fragment.

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Selected References

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  1. Acheson A., Edgar D., Timpl R., Thoenen H. Laminin increases both levels and activity of tyrosine hydroxylase in calf adrenal chromaffin cells. J Cell Biol. 1986 Jan;102(1):151–159. doi: 10.1083/jcb.102.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aumailley M., Timpl R. Attachment of cells to basement membrane collagen type IV. J Cell Biol. 1986 Oct;103(4):1569–1575. doi: 10.1083/jcb.103.4.1569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barsky S. H., Rao C. N., Hyams D., Liotta L. A. Characterization of a laminin receptor from human breast carcinoma tissue. Breast Cancer Res Treat. 1984;4(3):181–188. doi: 10.1007/BF01806483. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. 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;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Dessau W., Jilek F., Adelmann B. C., Hörmann H. Similarity of antigelatin factor and cold insoluble globulin. Biochim Biophys Acta. 1978 Mar 28;533(1):227–237. doi: 10.1016/0005-2795(78)90566-4. [DOI] [PubMed] [Google Scholar]
  6. Edgar D., Timpl R., Thoenen H. The heparin-binding domain of laminin is responsible for its effects on neurite outgrowth and neuronal survival. EMBO J. 1984 Jul;3(7):1463–1468. doi: 10.1002/j.1460-2075.1984.tb01997.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goldberg B. Binding of soluble type I collagen molecules to the fibroblast plasma membrane. Cell. 1979 Feb;16(2):265–275. doi: 10.1016/0092-8674(79)90004-7. [DOI] [PubMed] [Google Scholar]
  8. Goodman S. L., Newgreen D. Do cells show an inverse locomotory response to fibronectin and laminin substrates? EMBO J. 1985 Nov;4(11):2769–2771. doi: 10.1002/j.1460-2075.1985.tb04002.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Greenburg G., Hay E. D. Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells. J Cell Biol. 1982 Oct;95(1):333–339. doi: 10.1083/jcb.95.1.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horwitz A., Duggan K., Buck C., Beckerle M. C., Burridge K. Interaction of plasma membrane fibronectin receptor with talin--a transmembrane linkage. Nature. 1986 Apr 10;320(6062):531–533. doi: 10.1038/320531a0. [DOI] [PubMed] [Google Scholar]
  11. Horwitz A., Duggan K., Greggs R., Decker C., Buck C. The cell substrate attachment (CSAT) antigen has properties of a receptor for laminin and fibronectin. J Cell Biol. 1985 Dec;101(6):2134–2144. doi: 10.1083/jcb.101.6.2134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hubbard A. L., Cohn Z. A. Externally disposed plasma membrane proteins. I. Enzymatic iodination of mouse L cells. J Cell Biol. 1975 Feb;64(2):438–460. doi: 10.1083/jcb.64.2.438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johansson S., Kjellén L., Hök M., Timpl R. Substrate adhesion of rat hepatocytes: a comparison of laminin and fibronectin as attachment proteins. J Cell Biol. 1981 Jul;90(1):260–264. doi: 10.1083/jcb.90.1.260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kleinman H. K., Cannon F. B., Laurie G. W., Hassell J. R., Aumailley M., Terranova V. P., Martin G. R., DuBois-Dalcq M. Biological activities of laminin. J Cell Biochem. 1985;27(4):317–325. doi: 10.1002/jcb.240270402. [DOI] [PubMed] [Google Scholar]
  15. Kühl U., Ocalan M., Timpl R., von der Mark K. Role of laminin and fibronectin in selecting myogenic versus fibrogenic cells from skeletal muscle cells in vitro. Dev Biol. 1986 Oct;117(2):628–635. doi: 10.1016/0012-1606(86)90331-3. [DOI] [PubMed] [Google Scholar]
  16. Kühl U., Timpl R., von der Mark K. Synthesis of type IV collagen and laminin in cultures of skeletal muscle cells and their assembly on the surface of myotubes. Dev Biol. 1982 Oct;93(2):344–354. doi: 10.1016/0012-1606(82)90122-1. [DOI] [PubMed] [Google Scholar]
  17. Kühn K., Wiedemann H., Timpl R., Risteli J., Dieringer H., Voss T., Glanville R. W. Macromolecular structure of basement membrane collagens. FEBS Lett. 1981 Mar 9;125(1):123–128. doi: 10.1016/0014-5793(81)81012-5. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Landegren U. Measurement of cell numbers by means of the endogenous enzyme hexosaminidase. Applications to detection of lymphokines and cell surface antigens. J Immunol Methods. 1984 Mar 16;67(2):379–388. doi: 10.1016/0022-1759(84)90477-0. [DOI] [PubMed] [Google Scholar]
  20. Lane E. B., Goodman S. L., Trejdosiewicz L. K. Disruption of the keratin filament network during epithelial cell division. EMBO J. 1982;1(11):1365–1372. doi: 10.1002/j.1460-2075.1982.tb01324.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mauchamp J., Margotat A., Chambard M., Charrier B., Remy L., Michel-Bechet M. Polarity of three-dimensional structures derived from isolated hog thyroid cells in primary culture. Cell Tissue Res. 1979;204(3):417–430. doi: 10.1007/BF00233653. [DOI] [PubMed] [Google Scholar]
  22. Mollenhauer J., von der Mark K. Isolation and characterization of a collagen-binding glycoprotein from chondrocyte membranes. EMBO J. 1983;2(1):45–50. doi: 10.1002/j.1460-2075.1983.tb01378.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Newgreen D., Thiery J. P. Fibronectin in early avian embryos: synthesis and distribution along the migration pathways of neural crest cells. Cell Tissue Res. 1980;211(2):269–291. doi: 10.1007/BF00236449. [DOI] [PubMed] [Google Scholar]
  24. Ott U., Odermatt E., Engel J., Furthmayr H., Timpl R. Protease resistance and conformation of laminin. Eur J Biochem. 1982 Mar;123(1):63–72. doi: 10.1111/j.1432-1033.1982.tb06499.x. [DOI] [PubMed] [Google Scholar]
  25. Paulsson M., Deutzmann R., Timpl R., Dalzoppo D., Odermatt E., Engel J. Evidence for coiled-coil alpha-helical regions in the long arm of laminin. EMBO J. 1985 Feb;4(2):309–316. doi: 10.1002/j.1460-2075.1985.tb03630.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pierschbacher M. D., Ruoslahti E. Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature. 1984 May 3;309(5963):30–33. doi: 10.1038/309030a0. [DOI] [PubMed] [Google Scholar]
  27. Pierschbacher M. D., Ruoslahti E., Sundelin J., Lind P., Peterson P. A. The cell attachment domain of fibronectin. Determination of the primary structure. J Biol Chem. 1982 Aug 25;257(16):9593–9597. [PubMed] [Google Scholar]
  28. Pytela R., Pierschbacher M. D., Ruoslahti E. Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor. Cell. 1985 Jan;40(1):191–198. doi: 10.1016/0092-8674(85)90322-8. [DOI] [PubMed] [Google Scholar]
  29. Rao C. N., Margulies I. M., Tralka T. S., Terranova V. P., Madri J. A., Liotta L. A. Isolation of a subunit of laminin and its role in molecular structure and tumor cell attachment. J Biol Chem. 1982 Aug 25;257(16):9740–9744. [PubMed] [Google Scholar]
  30. Ruoslahti E., Hayman E. G., Pierschbacher M. D. Extracellular matrices and cell adhesion. Arteriosclerosis. 1985 Nov-Dec;5(6):581–594. doi: 10.1161/01.atv.5.6.581. [DOI] [PubMed] [Google Scholar]
  31. Sugrue S. P., Hay E. D. Response of basal epithelial cell surface and Cytoskeleton to solubilized extracellular matrix molecules. J Cell Biol. 1981 Oct;91(1):45–54. doi: 10.1083/jcb.91.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Terranova V. P., Rao C. N., Kalebic T., Margulies I. M., Liotta L. A. Laminin receptor on human breast carcinoma cells. Proc Natl Acad Sci U S A. 1983 Jan;80(2):444–448. doi: 10.1073/pnas.80.2.444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Timpl R., Dziadek M. Structure, development, and molecular pathology of basement membranes. Int Rev Exp Pathol. 1986;29:1–112. [PubMed] [Google Scholar]
  34. Timpl R., Johansson S., van Delden V., Oberbäumer I., Hök M. Characterization of protease-resistant fragments of laminin mediating attachment and spreading of rat hepatocytes. J Biol Chem. 1983 Jul 25;258(14):8922–8927. [PubMed] [Google Scholar]
  35. Timpl R., Rohde H., Robey P. G., Rennard S. I., Foidart J. M., Martin G. R. Laminin--a glycoprotein from basement membranes. J Biol Chem. 1979 Oct 10;254(19):9933–9937. [PubMed] [Google Scholar]
  36. von der Mark K., Kühl U. Laminin and its receptor. Biochim Biophys Acta. 1985 Dec 17;823(2):147–160. doi: 10.1016/0304-419x(85)90010-1. [DOI] [PubMed] [Google Scholar]

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