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. 1996 Oct 2;135(2):487–496. doi: 10.1083/jcb.135.2.487

Lateral dimerization is required for the homophilic binding activity of C-cadherin

PMCID: PMC2121050  PMID: 8896604

Abstract

Regulation of cadherin-mediated adhesion can occur rapidly at the cell surface. To understand the mechanism underlying cadherin regulation, it is essential to elucidate the homophilic binding mechanism that underlies all cadherin-mediated functions. Therefore, we have investigated the structural and functional properties of the extracellular segment of Xenopus C-cadherin using a purified, recombinant protein (CEC 1-5). CEC 1-5 supported adhesion of CHO cells expressing C-cadherin. The extracellular segment was also capable of mediating aggregation of microspheres. Chemical cross-linking and gel filtration revealed that CEC 1-5 formed dimers in the presence as well as absence of calcium. Analysis of the functional activity of purified dimers and monomers demonstrated that dimers retained substantially greater homophilic binding activity than monomers. These results demonstrate that lateral dimerization is necessary for homophilic binding between cadherin extracellular segments and suggest multiple potential mechanisms for the regulation of cadherin activity. Since the extracellular segment alone possessed significant homophilic binding activity, the adhesive activity of the extracellular segment in a cellular context was analyzed. The adhesion of CHO cells expressing a truncated version of C-cadherin lacking the cytoplasmic tail was compared to cells expressing the wild-type C-cadherin using a laminar flow assay on substrates coated with CEC 1-5. CHO cells expressing the truncated C-cadherin were able to attach to CEC 1-5 and to resist detachment by low shear forces, demonstrating that tailless C-cadherin can mediate basic, weak adhesion of CHO cells. However, cells expressing the truncated C-cadherin did not exhibit the complete adhesive activity of cells expressing wild-type C-cadherin. Cells expressing wild-type C-cadherin remained attached to CEC 1-5 at high shear forces, while cells expressing the tailless C-cadherin did not adhere well at high shear forces. These results suggest that there may be two states of cadherin-mediated adhesion. The first, relatively weak state can be mediated through interactions between the extracellular segments alone. The second strong adhesive state is critically dependent on the cytoplasmic tail.

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Selected References

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  1. Behrens J., Vakaet L., Friis R., Winterhager E., Van Roy F., Mareel M. M., Birchmeier W. Loss of epithelial differentiation and gain of invasiveness correlates with tyrosine phosphorylation of the E-cadherin/beta-catenin complex in cells transformed with a temperature-sensitive v-SRC gene. J Cell Biol. 1993 Feb;120(3):757–766. doi: 10.1083/jcb.120.3.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berlin C., Bargatze R. F., Campbell J. J., von Andrian U. H., Szabo M. C., Hasslen S. R., Nelson R. D., Berg E. L., Erlandsen S. L., Butcher E. C. alpha 4 integrins mediate lymphocyte attachment and rolling under physiologic flow. Cell. 1995 Feb 10;80(3):413–422. doi: 10.1016/0092-8674(95)90491-3. [DOI] [PubMed] [Google Scholar]
  3. Bixby J. L., Zhang R. Purified N-cadherin is a potent substrate for the rapid induction of neurite outgrowth. J Cell Biol. 1990 Apr;110(4):1253–1260. doi: 10.1083/jcb.110.4.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brieher W. M., Gumbiner B. M. Regulation of C-cadherin function during activin induced morphogenesis of Xenopus animal caps. J Cell Biol. 1994 Jul;126(2):519–527. doi: 10.1083/jcb.126.2.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davis S. J., Ward H. A., Puklavec M. J., Willis A. C., Williams A. F., Barclay A. N. High level expression in Chinese hamster ovary cells of soluble forms of CD4 T lymphocyte glycoprotein including glycosylation variants. J Biol Chem. 1990 Jun 25;265(18):10410–10418. [PubMed] [Google Scholar]
  6. Fleming T. P., Johnson M. H. From egg to epithelium. Annu Rev Cell Biol. 1988;4:459–485. doi: 10.1146/annurev.cb.04.110188.002331. [DOI] [PubMed] [Google Scholar]
  7. Fujimori T., Takeichi M. Disruption of epithelial cell-cell adhesion by exogenous expression of a mutated nonfunctional N-cadherin. Mol Biol Cell. 1993 Jan;4(1):37–47. doi: 10.1091/mbc.4.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grumet M., Friedlander D. R., Edelman G. M. Evidence for the binding of Ng-CAM to laminin. Cell Adhes Commun. 1993 Sep;1(2):177–190. doi: 10.3109/15419069309095693. [DOI] [PubMed] [Google Scholar]
  9. Gumbiner B. M. Cell adhesion: the molecular basis of tissue architecture and morphogenesis. Cell. 1996 Feb 9;84(3):345–357. doi: 10.1016/s0092-8674(00)81279-9. [DOI] [PubMed] [Google Scholar]
  10. Hamaguchi M., Matsuyoshi N., Ohnishi Y., Gotoh B., Takeichi M., Nagai Y. p60v-src causes tyrosine phosphorylation and inactivation of the N-cadherin-catenin cell adhesion system. EMBO J. 1993 Jan;12(1):307–314. doi: 10.1002/j.1460-2075.1993.tb05658.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Herrenknecht K., Kemler R. Characterization of recombinant E-cadherin (uvomorulin) expressed in insect cells. J Cell Sci Suppl. 1993;17:147–154. doi: 10.1242/jcs.1993.supplement_17.21. [DOI] [PubMed] [Google Scholar]
  12. Hirano S., Kimoto N., Shimoyama Y., Hirohashi S., Takeichi M. Identification of a neural alpha-catenin as a key regulator of cadherin function and multicellular organization. Cell. 1992 Jul 24;70(2):293–301. doi: 10.1016/0092-8674(92)90103-j. [DOI] [PubMed] [Google Scholar]
  13. Lee C. H., Gumbiner B. M. Disruption of gastrulation movements in Xenopus by a dominant-negative mutant for C-cadherin. Dev Biol. 1995 Oct;171(2):363–373. doi: 10.1006/dbio.1995.1288. [DOI] [PubMed] [Google Scholar]
  14. Levine E., Lee C. H., Kintner C., Gumbiner B. M. Selective disruption of E-cadherin function in early Xenopus embryos by a dominant negative mutant. Development. 1994 Apr;120(4):901–909. doi: 10.1242/dev.120.4.901. [DOI] [PubMed] [Google Scholar]
  15. Nagafuchi A., Takeichi M. Cell binding function of E-cadherin is regulated by the cytoplasmic domain. EMBO J. 1988 Dec 1;7(12):3679–3684. doi: 10.1002/j.1460-2075.1988.tb03249.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nagar B., Overduin M., Ikura M., Rini J. M. Structural basis of calcium-induced E-cadherin rigidification and dimerization. Nature. 1996 Mar 28;380(6572):360–364. doi: 10.1038/380360a0. [DOI] [PubMed] [Google Scholar]
  17. Ozawa M., Baribault H., Kemler R. The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species. EMBO J. 1989 Jun;8(6):1711–1717. doi: 10.1002/j.1460-2075.1989.tb03563.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ozawa M., Ringwald M., Kemler R. Uvomorulin-catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4246–4250. doi: 10.1073/pnas.87.11.4246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Paradies N. E., Grunwald G. B. Purification and characterization of NCAD90, a soluble endogenous form of N-cadherin, which is generated by proteolysis during retinal development and retains adhesive and neurite-promoting function. J Neurosci Res. 1993 Sep 1;36(1):33–45. doi: 10.1002/jnr.490360105. [DOI] [PubMed] [Google Scholar]
  20. Payne H. R., Lemmon V. Glial cells of the O-2A lineage bind preferentially to N-cadherin and develop distinct morphologies. Dev Biol. 1993 Oct;159(2):595–607. doi: 10.1006/dbio.1993.1267. [DOI] [PubMed] [Google Scholar]
  21. Pokutta S., Herrenknecht K., Kemler R., Engel J. Conformational changes of the recombinant extracellular domain of E-cadherin upon calcium binding. Eur J Biochem. 1994 Aug 1;223(3):1019–1026. doi: 10.1111/j.1432-1033.1994.tb19080.x. [DOI] [PubMed] [Google Scholar]
  22. Schwartz M. A., Schaller M. D., Ginsberg M. H. Integrins: emerging paradigms of signal transduction. Annu Rev Cell Dev Biol. 1995;11:549–599. doi: 10.1146/annurev.cb.11.110195.003001. [DOI] [PubMed] [Google Scholar]
  23. Shapiro L., Fannon A. M., Kwong P. D., Thompson A., Lehmann M. S., Grübel G., Legrand J. F., Als-Nielsen J., Colman D. R., Hendrickson W. A. Structural basis of cell-cell adhesion by cadherins. Nature. 1995 Mar 23;374(6520):327–337. doi: 10.1038/374327a0. [DOI] [PubMed] [Google Scholar]
  24. Wheelock M. J., Buck C. A., Bechtol K. B., Damsky C. H. Soluble 80-kd fragment of cell-CAM 120/80 disrupts cell-cell adhesion. J Cell Biochem. 1987 Jul;34(3):187–202. doi: 10.1002/jcb.240340305. [DOI] [PubMed] [Google Scholar]
  25. Williams C. L., Hayes V. Y., Hummel A. M., Tarara J. E., Halsey T. J. Regulation of E-cadherin-mediated adhesion by muscarinic acetylcholine receptors in small cell lung carcinoma. J Cell Biol. 1993 May;121(3):643–654. doi: 10.1083/jcb.121.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]

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