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. 1995 Aug 1;309(Pt 3):765–771. doi: 10.1042/bj3090765

Evidence for the location of a binding sequence for the alpha 2 beta 1 integrin of endothelial cells, in the beta 1 subunit of laminin.

P A Underwood 1, F A Bennett 1, A Kirkpatrick 1, P A Bean 1, B A Moss 1
PMCID: PMC1135698  PMID: 7639691

Abstract

To date no specific location on laminin 1 for the binding of alpha 2 beta 1 integrin has been described, although recent evidence supports a location in the E1XNd fragment of the cross region. We have identified a peptide sequence from this region, in the beta 1 chain of laminin 1, YGYYGDALR, which inhibits the adhesion of endothelial cells to laminin 1 and type-IV collagen. A structurally related sequence from the CNBr-cleaved fragment CB3 of the alpha 1 chain of collagen type IV, FYFDLR, inhibits endothelial cell adhesion to both collagen types I and IV and laminin 1. The CB3 fragment containing the FYFDLR sequence has been shown to contain binding sites for both alpha 1 beta 1 and alpha 2 beta 1 integrins. Present experiments with anti-integrin antibodies indicate that the alpha 2 beta 1 integrin on endothelial cells can account for all the cell binding to collagen types I and IV, and that this integrin makes a major contribution towards the adhesion of these cells to laminin 1. We therefore propose that the peptide FYFDLR participates in alpha 2 beta 1 binding to collagen type IV and that the putatively structurally similar peptide, YGYYGDALR, participates in alpha 2 beta 1 binding to laminin 1. This is the first account of structurally related peptide sequences from laminin 1 and type-IV collagen which show reciprocal inhibition of cell adhesion to either ligand and which might form part of a common integrin-binding site, as well as the first suggestion of a precise location contributing to the alpha 2 beta 1 integrin binding site on laminin 1.

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Selected References

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

  1. Akiyama S. K., Nagata K., Yamada K. M. Cell surface receptors for extracellular matrix components. Biochim Biophys Acta. 1990 Feb 28;1031(1):91–110. doi: 10.1016/0304-4157(90)90004-v. [DOI] [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. Aumailley M., Timpl R., Sonnenberg A. Antibody to integrin alpha 6 subunit specifically inhibits cell-binding to laminin fragment 8. Exp Cell Res. 1990 May;188(1):55–60. doi: 10.1016/0014-4827(90)90277-h. [DOI] [PubMed] [Google Scholar]
  4. Brazel D., Pollner R., Oberbäumer I., Kühn K. Human basement membrane collagen (type IV). The amino acid sequence of the alpha 2(IV) chain and its comparison with the alpha 1(IV) chain reveals deletions in the alpha 1(IV) chain. Eur J Biochem. 1988 Feb 15;172(1):35–42. doi: 10.1111/j.1432-1033.1988.tb13852.x. [DOI] [PubMed] [Google Scholar]
  5. Burgeson R. E., Chiquet M., Deutzmann R., Ekblom P., Engel J., Kleinman H., Martin G. R., Meneguzzi G., Paulsson M., Sanes J. A new nomenclature for the laminins. Matrix Biol. 1994 Apr;14(3):209–211. doi: 10.1016/0945-053x(94)90184-8. [DOI] [PubMed] [Google Scholar]
  6. Chan B. M., Hemler M. E. Multiple functional forms of the integrin VLA-2 can be derived from a single alpha 2 cDNA clone: interconversion of forms induced by an anti-beta 1 antibody. J Cell Biol. 1993 Jan;120(2):537–543. doi: 10.1083/jcb.120.2.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Davis G. E. Affinity of integrins for damaged extracellular matrix: alpha v beta 3 binds to denatured collagen type I through RGD sites. Biochem Biophys Res Commun. 1992 Feb 14;182(3):1025–1031. doi: 10.1016/0006-291x(92)91834-d. [DOI] [PubMed] [Google Scholar]
  8. Dedhar S., Ruoslahti E., Pierschbacher M. D. A cell surface receptor complex for collagen type I recognizes the Arg-Gly-Asp sequence. J Cell Biol. 1987 Mar;104(3):585–593. doi: 10.1083/jcb.104.3.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elices M. J., Hemler M. E. The human integrin VLA-2 is a collagen receptor on some cells and a collagen/laminin receptor on others. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9906–9910. doi: 10.1073/pnas.86.24.9906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gehlsen K. R., Dickerson K., Argraves W. S., Engvall E., Ruoslahti E. Subunit structure of a laminin-binding integrin and localization of its binding site on laminin. J Biol Chem. 1989 Nov 15;264(32):19034–19038. [PubMed] [Google Scholar]
  11. Gehlsen K. R., Sriramarao P., Furcht L. T., Skubitz A. P. A synthetic peptide derived from the carboxy terminus of the laminin A chain represents a binding site for the alpha 3 beta 1 integrin. J Cell Biol. 1992 Apr;117(2):449–459. doi: 10.1083/jcb.117.2.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Giltay J. C., Brinkman H. J., Modderman P. W., von dem Borne A. E., van Mourik J. A. Human vascular endothelial cells express a membrane protein complex immunochemically indistinguishable from the platelet VLA-2 (glycoprotein Ia-IIa) complex. Blood. 1989 Apr;73(5):1235–1241. [PubMed] [Google Scholar]
  13. Glanville R. W., Breitkreutz D., Meitinger M., Fietzek P. P. Completion of the amino acid sequence of the alpha 1 chain from type I calf skin collagen. Amino acid sequence of alpha 1(I)B8. Biochem J. 1983 Oct 1;215(1):183–189. doi: 10.1042/bj2150183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Graf J., Iwamoto Y., Sasaki M., Martin G. R., Kleinman H. K., Robey F. A., Yamada Y. Identification of an amino acid sequence in laminin mediating cell attachment, chemotaxis, and receptor binding. Cell. 1987 Mar 27;48(6):989–996. doi: 10.1016/0092-8674(87)90707-0. [DOI] [PubMed] [Google Scholar]
  15. Graf J., Ogle R. C., Robey F. A., Sasaki M., Martin G. R., Yamada Y., Kleinman H. K. A pentapeptide from the laminin B1 chain mediates cell adhesion and binds the 67,000 laminin receptor. Biochemistry. 1987 Nov 3;26(22):6896–6900. doi: 10.1021/bi00396a004. [DOI] [PubMed] [Google Scholar]
  16. Grant D. S., Tashiro K., Segui-Real B., Yamada Y., Martin G. R., Kleinman H. K. Two different laminin domains mediate the differentiation of human endothelial cells into capillary-like structures in vitro. Cell. 1989 Sep 8;58(5):933–943. doi: 10.1016/0092-8674(89)90945-8. [DOI] [PubMed] [Google Scholar]
  17. Gray A., Dull T. J., Ullrich A. Nucleotide sequence of epidermal growth factor cDNA predicts a 128,000-molecular weight protein precursor. Nature. 1983 Jun 23;303(5919):722–725. doi: 10.1038/303722a0. [DOI] [PubMed] [Google Scholar]
  18. Gullberg D., Gehlsen K. R., Turner D. C., Ahlén K., Zijenah L. S., Barnes M. J., Rubin K. Analysis of alpha 1 beta 1, alpha 2 beta 1 and alpha 3 beta 1 integrins in cell--collagen interactions: identification of conformation dependent alpha 1 beta 1 binding sites in collagen type I. EMBO J. 1992 Nov;11(11):3865–3873. doi: 10.1002/j.1460-2075.1992.tb05479.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kern A., Eble J., Golbik R., Kühn K. Interaction of type IV collagen with the isolated integrins alpha 1 beta 1 and alpha 2 beta 1. Eur J Biochem. 1993 Jul 1;215(1):151–159. doi: 10.1111/j.1432-1033.1993.tb18017.x. [DOI] [PubMed] [Google Scholar]
  21. Kirchhofer D., Languino L. R., Ruoslahti E., Pierschbacher M. D. Alpha 2 beta 1 integrins from different cell types show different binding specificities. J Biol Chem. 1990 Jan 15;265(2):615–618. [PubMed] [Google Scholar]
  22. Koliakos G. G., Kouzi-Koliakos K., Furcht L. T., Reger L. A., Tsilibary E. C. The binding of heparin to type IV collagen: domain specificity with identification of peptide sequences from the alpha 1(IV) and alpha 2(IV) which preferentially bind heparin. J Biol Chem. 1989 Feb 5;264(4):2313–2323. [PubMed] [Google Scholar]
  23. König W., Geiger R. Eine neue Methode zur Synthese von Peptiden: Aktivierung der Carboxylgruppe mit Dicyclohexycarbodiimid unter Zusatz von 1-Hydroxy-benzotriazolen. Chem Ber. 1970;103(3):788–798. doi: 10.1002/cber.19701030319. [DOI] [PubMed] [Google Scholar]
  24. Lampugnani M. G., Resnati M., Dejana E., Marchisio P. C. The role of integrins in the maintenance of endothelial monolayer integrity. J Cell Biol. 1991 Feb;112(3):479–490. doi: 10.1083/jcb.112.3.479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Languino L. R., Gehlsen K. R., Wayner E., Carter W. G., Engvall E., Ruoslahti E. Endothelial cells use alpha 2 beta 1 integrin as a laminin receptor. J Cell Biol. 1989 Nov;109(5):2455–2462. doi: 10.1083/jcb.109.5.2455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Maciag T., Cerundolo J., Ilsley S., Kelley P. R., Forand R. An endothelial cell growth factor from bovine hypothalamus: identification and partial characterization. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5674–5678. doi: 10.1073/pnas.76.11.5674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. McAuslan B. R., Reilly W., Hannan G. N. Stimulation of endothelia cell proliferation by precursors of thymidylate. J Cell Physiol. 1979 Jul;100(1):87–93. doi: 10.1002/jcp.1041000109. [DOI] [PubMed] [Google Scholar]
  28. McCarthy J. B., Skubitz A. P., Qi Z., Yi X. Y., Mickelson D. J., Klein D. J., Furcht L. T. RGD-independent cell adhesion to the carboxy-terminal heparin-binding fragment of fibronectin involves heparin-dependent and -independent activities. J Cell Biol. 1990 Mar;110(3):777–787. doi: 10.1083/jcb.110.3.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nissinen M., Vuolteenaho R., Boot-Handford R., Kallunki P., Tryggvason K. Primary structure of the human laminin A chain. Limited expression in human tissues. Biochem J. 1991 Jun 1;276(Pt 2):369–379. doi: 10.1042/bj2760369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. O'Connell P. J., Faull R., Russ G. R., D'Apice A. J. VLA-2 is a collagen receptor on endothelial cells. Immunol Cell Biol. 1991 Apr;69(Pt 2):103–110. doi: 10.1038/icb.1991.16. [DOI] [PubMed] [Google Scholar]
  31. Oliver M. H., Harrison N. K., Bishop J. E., Cole P. J., Laurent G. J. A rapid and convenient assay for counting cells cultured in microwell plates: application for assessment of growth factors. J Cell Sci. 1989 Mar;92(Pt 3):513–518. doi: 10.1242/jcs.92.3.513. [DOI] [PubMed] [Google Scholar]
  32. Pfaff M., Göhring W., Brown J. C., Timpl R. Binding of purified collagen receptors (alpha 1 beta 1, alpha 2 beta 1) and RGD-dependent integrins to laminins and laminin fragments. Eur J Biochem. 1994 Nov 1;225(3):975–984. doi: 10.1111/j.1432-1033.1994.0975b.x. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Price M. R., Sekowski M., Hooi D. S., Durrant L. G., Hudecz F., Tendler S. J. Measurement of antibody binding to antigenic peptides conjugated in situ to albumin-coated microtitre plates. J Immunol Methods. 1993 Feb 26;159(1-2):277–281. doi: 10.1016/0022-1759(93)90168-7. [DOI] [PubMed] [Google Scholar]
  35. Ruoslahti E., Hayman E. G., Pierschbacher M., Engvall E. Fibronectin: purification, immunochemical properties, and biological activities. Methods Enzymol. 1982;82(Pt A):803–831. doi: 10.1016/0076-6879(82)82103-4. [DOI] [PubMed] [Google Scholar]
  36. Ruoslahti E., Pierschbacher M. D. Arg-Gly-Asp: a versatile cell recognition signal. Cell. 1986 Feb 28;44(4):517–518. doi: 10.1016/0092-8674(86)90259-x. [DOI] [PubMed] [Google Scholar]
  37. Sasaki M., Kato S., Kohno K., Martin G. R., Yamada Y. Sequence of the cDNA encoding the laminin B1 chain reveals a multidomain protein containing cysteine-rich repeats. Proc Natl Acad Sci U S A. 1987 Feb;84(4):935–939. doi: 10.1073/pnas.84.4.935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Skubitz A. P., Letourneau P. C., Wayner E., Furcht L. T. Synthetic peptides from the carboxy-terminal globular domain of the A chain of laminin: their ability to promote cell adhesion and neurite outgrowth, and interact with heparin and the beta 1 integrin subunit. J Cell Biol. 1991 Nov;115(4):1137–1148. doi: 10.1083/jcb.115.4.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sonnenberg A., Linders C. J., Modderman P. W., Damsky C. H., Aumailley M., Timpl R. Integrin recognition of different cell-binding fragments of laminin (P1, E3, E8) and evidence that alpha 6 beta 1 but not alpha 6 beta 4 functions as a major receptor for fragment E8. J Cell Biol. 1990 Jun;110(6):2145–2155. doi: 10.1083/jcb.110.6.2145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Staatz W. D., Fok K. F., Zutter M. M., Adams S. P., Rodriguez B. A., Santoro S. A. Identification of a tetrapeptide recognition sequence for the alpha 2 beta 1 integrin in collagen. J Biol Chem. 1991 Apr 25;266(12):7363–7367. [PubMed] [Google Scholar]
  41. Tashiro K., Sephel G. C., Greatorex D., Sasaki M., Shirashi N., Martin G. R., Kleinman H. K., Yamada Y. The RGD containing site of the mouse laminin A chain is active for cell attachment, spreading, migration and neurite outgrowth. J Cell Physiol. 1991 Mar;146(3):451–459. doi: 10.1002/jcp.1041460316. [DOI] [PubMed] [Google Scholar]
  42. Underwood P. A., Bennett F. A. A comparison of the biological activities of the cell-adhesive proteins vitronectin and fibronectin. J Cell Sci. 1989 Aug;93(Pt 4):641–649. doi: 10.1242/jcs.93.4.641. [DOI] [PubMed] [Google Scholar]
  43. Underwood P. A., Steele J. G., Dalton B. A., Bennett F. A. Solid-phase monoclonal antibodies. A novel method of directing the function of biologically active molecules by presenting a specific orientation. J Immunol Methods. 1990 Feb 20;127(1):91–101. doi: 10.1016/0022-1759(90)90344-u. [DOI] [PubMed] [Google Scholar]
  44. Vandenberg P., Kern A., Ries A., Luckenbill-Edds L., Mann K., Kühn K. Characterization of a type IV collagen major cell binding site with affinity to the alpha 1 beta 1 and the alpha 2 beta 1 integrins. J Cell Biol. 1991 Jun;113(6):1475–1483. doi: 10.1083/jcb.113.6.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Yamada K. M. Adhesive recognition sequences. J Biol Chem. 1991 Jul 15;266(20):12809–12812. [PubMed] [Google Scholar]

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