Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1999 Mar 1;338(Pt 2):471–478.

Dephosphorylation of the catenins p120 and p100 in endothelial cells in response to inflammatory stimuli.

M J Ratcliffe 1, C Smales 1, J M Staddon 1
PMCID: PMC1220075  PMID: 10024525

Abstract

Inflammatory mediators such as histamine and thrombin increase the tight-junction permeability of endothelial cells. Tight-junction permeability may be independently controlled, but is dependent on the adherens junction, where adhesion is achieved through homotypic interaction of cadherins, which in turn are associated with cytoplasmic proteins, the catenins. p120, also termed p120(cas)/p120(ctn), and its splice variant, p100, are catenins. p120, originally discovered as a substrate of the tyrosine kinase Src, is also a target for a protein kinase C-stimulated pathway in epithelial cells, causing its serine/threonine dephosphorylation. The present study shows that pharmacological activation of protein kinase C stimulated a similar pathway in endothelial cells. Activation of receptors for agents such as histamine (H1), thrombin and lysophosphatidic acid in the endothelial cells also caused serine/threonine dephosphorylation of p120 and p100, suggesting physiological relevance. However, protein kinase C inhibitors, although blocking the effect of pharmacological activation of protein kinase C, did not block the effects due to receptor activation. Calcium mobilization and the myosin-light-chain-kinase pathway do not participate in p120/p100 signalling. In conclusion, endothelial cells possess protein kinase C-dependent and -independent pathways regulating p120/p100 serine/threonine phosphorylation. These data describe a new connection between inflammatory agents, receptor-stimulated signalling and pathways potentially influencing intercellular adhesion in endothelial cells.

Full Text

The Full Text of this article is available as a PDF (237.3 KB).

Selected References

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

  1. Aberle H., Butz S., Stappert J., Weissig H., Kemler R., Hoschuetzky H. Assembly of the cadherin-catenin complex in vitro with recombinant proteins. J Cell Sci. 1994 Dec;107(Pt 12):3655–3663. doi: 10.1242/jcs.107.12.3655. [DOI] [PubMed] [Google Scholar]
  2. Aberle H., Schwartz H., Kemler R. Cadherin-catenin complex: protein interactions and their implications for cadherin function. J Cell Biochem. 1996 Jun 15;61(4):514–523. doi: 10.1002/(SICI)1097-4644(19960616)61:4%3C514::AID-JCB4%3E3.0.CO;2-R. [DOI] [PubMed] [Google Scholar]
  3. Birchmeier W., Behrens J. Cadherin expression in carcinomas: role in the formation of cell junctions and the prevention of invasiveness. Biochim Biophys Acta. 1994 May 27;1198(1):11–26. doi: 10.1016/0304-419x(94)90003-5. [DOI] [PubMed] [Google Scholar]
  4. Boyle W. J., van der Geer P., Hunter T. Phosphopeptide mapping and phosphoamino acid analysis by two-dimensional separation on thin-layer cellulose plates. Methods Enzymol. 1991;201:110–149. doi: 10.1016/0076-6879(91)01013-r. [DOI] [PubMed] [Google Scholar]
  5. Carson M. R., Shasby S. S., Shasby D. M. Histamine and inositol phosphate accumulation in endothelium: cAMP and a G protein. Am J Physiol. 1989 Oct;257(4 Pt 1):L259–L264. doi: 10.1152/ajplung.1989.257.4.L259. [DOI] [PubMed] [Google Scholar]
  6. Daniel J. M., Reynolds A. B. Tyrosine phosphorylation and cadherin/catenin function. Bioessays. 1997 Oct;19(10):883–891. doi: 10.1002/bies.950191008. [DOI] [PubMed] [Google Scholar]
  7. Edgell C. J., McDonald C. C., Graham J. B. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3734–3737. doi: 10.1073/pnas.80.12.3734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goeckeler Z. M., Wysolmerski R. B. Myosin light chain kinase-regulated endothelial cell contraction: the relationship between isometric tension, actin polymerization, and myosin phosphorylation. J Cell Biol. 1995 Aug;130(3):613–627. doi: 10.1083/jcb.130.3.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  10. 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]
  11. Haselton F. R., Heimark R. L. Role of cadherins 5 and 13 in the aortic endothelial barrier. J Cell Physiol. 1997 Jun;171(3):243–251. doi: 10.1002/(SICI)1097-4652(199706)171:3<243::AID-JCP2>3.0.CO;2-O. [DOI] [PubMed] [Google Scholar]
  12. Hidaka H., Inagaki M., Kawamoto S., Sasaki Y. Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry. 1984 Oct 9;23(21):5036–5041. doi: 10.1021/bi00316a032. [DOI] [PubMed] [Google Scholar]
  13. Huang A. J., Manning J. E., Bandak T. M., Ratau M. C., Hanser K. R., Silverstein S. C. Endothelial cell cytosolic free calcium regulates neutrophil migration across monolayers of endothelial cells. J Cell Biol. 1993 Mar;120(6):1371–1380. doi: 10.1083/jcb.120.6.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jacobson B. C., Pober J. S., Fenton J. W., 2nd, Ewenstein B. M. Thrombin and histamine rapidly stimulate the phosphorylation of the myristoylated alanine-rich C-kinase substrate in human umbilical vein endothelial cells: evidence for distinct patterns of protein kinase activation. J Cell Physiol. 1992 Jul;152(1):166–176. doi: 10.1002/jcp.1041520121. [DOI] [PubMed] [Google Scholar]
  15. Kimura K., Ito M., Amano M., Chihara K., Fukata Y., Nakafuku M., Yamamori B., Feng J., Nakano T., Okawa K. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase) Science. 1996 Jul 12;273(5272):245–248. doi: 10.1126/science.273.5272.245. [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. Lampugnani M. G., Dejana E. Interendothelial junctions: structure, signalling and functional roles. Curr Opin Cell Biol. 1997 Oct;9(5):674–682. doi: 10.1016/s0955-0674(97)80121-4. [DOI] [PubMed] [Google Scholar]
  18. Levin E. G., Santell L. Thrombin- and histamine-induced signal transduction in human endothelial cells. Stimulation and agonist-dependent desensitization of protein phosphorylation. J Biol Chem. 1991 Jan 5;266(1):174–181. [PubMed] [Google Scholar]
  19. Linder M. E., Burr J. G. Nonmyristoylated p60v-src fails to phosphorylate proteins of 115-120 kDa in chicken embryo fibroblasts. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2608–2612. doi: 10.1073/pnas.85.8.2608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McCrea P. D., Turck C. W., Gumbiner B. A homolog of the armadillo protein in Drosophila (plakoglobin) associated with E-cadherin. Science. 1991 Nov 29;254(5036):1359–1361. doi: 10.1126/science.1962194. [DOI] [PubMed] [Google Scholar]
  21. Mellor H., Parker P. J. The extended protein kinase C superfamily. Biochem J. 1998 Jun 1;332(Pt 2):281–292. doi: 10.1042/bj3320281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mo Y. Y., Reynolds A. B. Identification of murine p120 isoforms and heterogeneous expression of p120cas isoforms in human tumor cell lines. Cancer Res. 1996 Jun 1;56(11):2633–2640. [PubMed] [Google Scholar]
  23. Moolenaar W. H., Kranenburg O., Postma F. R., Zondag G. C. Lysophosphatidic acid: G-protein signalling and cellular responses. Curr Opin Cell Biol. 1997 Apr;9(2):168–173. doi: 10.1016/s0955-0674(97)80059-2. [DOI] [PubMed] [Google Scholar]
  24. Moy A. B., Shasby S. S., Scott B. D., Shasby D. M. The effect of histamine and cyclic adenosine monophosphate on myosin light chain phosphorylation in human umbilical vein endothelial cells. J Clin Invest. 1993 Sep;92(3):1198–1206. doi: 10.1172/JCI116690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. 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]
  28. Parkos C. A. Molecular events in neutrophil transepithelial migration. Bioessays. 1997 Oct;19(10):865–873. doi: 10.1002/bies.950191006. [DOI] [PubMed] [Google Scholar]
  29. Peifer M., Berg S., Reynolds A. B. A repeating amino acid motif shared by proteins with diverse cellular roles. Cell. 1994 Mar 11;76(5):789–791. doi: 10.1016/0092-8674(94)90353-0. [DOI] [PubMed] [Google Scholar]
  30. Pollock W. K., Wreggett K. A., Irvine R. F. Inositol phosphate production and Ca2+ mobilization in human umbilical-vein endothelial cells stimulated by thrombin and histamine. Biochem J. 1988 Dec 1;256(2):371–376. doi: 10.1042/bj2560371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ratcliffe M. J., Rubin L. L., Staddon J. M. Dephosphorylation of the cadherin-associated p100/p120 proteins in response to activation of protein kinase C in epithelial cells. J Biol Chem. 1997 Dec 12;272(50):31894–31901. doi: 10.1074/jbc.272.50.31894. [DOI] [PubMed] [Google Scholar]
  32. Reynolds A. B., Daniel J., McCrea P. D., Wheelock M. J., Wu J., Zhang Z. Identification of a new catenin: the tyrosine kinase substrate p120cas associates with E-cadherin complexes. Mol Cell Biol. 1994 Dec;14(12):8333–8342. doi: 10.1128/mcb.14.12.8333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Reynolds A. B., Roesel D. J., Kanner S. B., Parsons J. T. Transformation-specific tyrosine phosphorylation of a novel cellular protein in chicken cells expressing oncogenic variants of the avian cellular src gene. Mol Cell Biol. 1989 Feb;9(2):629–638. doi: 10.1128/mcb.9.2.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rimm D. L., Koslov E. R., Kebriaei P., Cianci C. D., Morrow J. S. Alpha 1(E)-catenin is an actin-binding and -bundling protein mediating the attachment of F-actin to the membrane adhesion complex. Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8813–8817. doi: 10.1073/pnas.92.19.8813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rotrosen D., Gallin J. I. Histamine type I receptor occupancy increases endothelial cytosolic calcium, reduces F-actin, and promotes albumin diffusion across cultured endothelial monolayers. J Cell Biol. 1986 Dec;103(6 Pt 1):2379–2387. doi: 10.1083/jcb.103.6.2379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Shibamoto S., Hayakawa M., Takeuchi K., Hori T., Miyazawa K., Kitamura N., Johnson K. R., Wheelock M. J., Matsuyoshi N., Takeichi M. Association of p120, a tyrosine kinase substrate, with E-cadherin/catenin complexes. J Cell Biol. 1995 Mar;128(5):949–957. doi: 10.1083/jcb.128.5.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Staddon J. M., Herrenknecht K., Smales C., Rubin L. L. Evidence that tyrosine phosphorylation may increase tight junction permeability. J Cell Sci. 1995 Feb;108(Pt 2):609–619. doi: 10.1242/jcs.108.2.609. [DOI] [PubMed] [Google Scholar]
  38. Staddon J. M., Rubin L. L. Cell adhesion, cell junctions and the blood-brain barrier. Curr Opin Neurobiol. 1996 Oct;6(5):622–627. doi: 10.1016/s0959-4388(96)80094-8. [DOI] [PubMed] [Google Scholar]
  39. Staddon J. M., Smales C., Schulze C., Esch F. S., Rubin L. L. p120, a p120-related protein (p100), and the cadherin/catenin complex. J Cell Biol. 1995 Jul;130(2):369–381. doi: 10.1083/jcb.130.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Takeda H., Nagafuchi A., Yonemura S., Tsukita S., Behrens J., Birchmeier W., Tsukita S. V-src kinase shifts the cadherin-based cell adhesion from the strong to the weak state and beta catenin is not required for the shift. J Cell Biol. 1995 Dec;131(6 Pt 2):1839–1847. doi: 10.1083/jcb.131.6.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Thastrup O. Role of Ca2(+)-ATPases in regulation of cellular Ca2+ signalling, as studied with the selective microsomal Ca2(+)-ATPase inhibitor, thapsigargin. Agents Actions. 1990 Jan;29(1-2):8–15. doi: 10.1007/BF01964706. [DOI] [PubMed] [Google Scholar]
  42. Toullec D., Pianetti P., Coste H., Bellevergue P., Grand-Perret T., Ajakane M., Baudet V., Boissin P., Boursier E., Loriolle F. The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J Biol Chem. 1991 Aug 25;266(24):15771–15781. [PubMed] [Google Scholar]
  43. Wilkinson S. E., Parker P. J., Nixon J. S. Isoenzyme specificity of bisindolylmaleimides, selective inhibitors of protein kinase C. Biochem J. 1993 Sep 1;294(Pt 2):335–337. doi: 10.1042/bj2940335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Yap A. S., Niessen C. M., Gumbiner B. M. The juxtamembrane region of the cadherin cytoplasmic tail supports lateral clustering, adhesive strengthening, and interaction with p120ctn. J Cell Biol. 1998 May 4;141(3):779–789. doi: 10.1083/jcb.141.3.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Zhao Y., Davis H. W. Thrombin-induced phosphorylation of the myristoylated alanine-rich C kinase substrate (MARCKS) protein in bovine pulmonary artery endothelial cells. J Cell Physiol. 1996 Nov;169(2):350–357. doi: 10.1002/(SICI)1097-4652(199611)169:2<350::AID-JCP14>3.0.CO;2-D. [DOI] [PubMed] [Google Scholar]

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

RESOURCES