Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1998 Jan;74(1):514–522. doi: 10.1016/S0006-3495(98)77808-7

Membrane bending modulus and adhesion energy of wild-type and mutant cells of Dictyostelium lacking talin or cortexillins.

R Simson 1, E Wallraff 1, J Faix 1, J Niewöhner 1, G Gerisch 1, E Sackmann 1
PMCID: PMC1299403  PMID: 9449351

Abstract

We have employed an interferometric technique for the local measurement of bending modulus, membrane tension, and adhesion energy of motile cells adhering to a substrate. Wild-type and mutant cells of Dictyostelium discoideum were incubated in a flow chamber. The flow-induced deformation of a cell near its adhesion area was determined by quantitative reflection interference contrast microscopy (RICM) and analyzed in terms of the elastic boundary conditions: equilibrium of tensions and bending moments at the contact line. This technique was employed to quantify changes caused by the lack of talin, a protein that couples the actin network to the plasma membrane, or by the lack of cortexillin I or II, two isoforms of the actin-bundling protein cortexillin. Cells lacking either cortexillin I or II exhibited reduced bending moduli of 95 and 160 k(B)T, respectively, as compared to 390 k(B)T, obtained for wild-type cells. No significant difference was found for the adhesion energies of wild-type and cortexillin mutant cells. In cells lacking talin, not only a strongly reduced bending modulus of 70 k(B)T, but also a low adhesion energy one-fourth of that in wild-type cells was measured.

Full Text

The Full Text of this article is available as a PDF (305.5 KB).

Selected References

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

  1. Albigès-Rizo C., Frachet P., Block M. R. Down regulation of talin alters cell adhesion and the processing of the alpha 5 beta 1 integrin. J Cell Sci. 1995 Oct;108(Pt 10):3317–3329. doi: 10.1242/jcs.108.10.3317. [DOI] [PubMed] [Google Scholar]
  2. Alon R., Hammer D. A., Springer T. A. Lifetime of the P-selectin-carbohydrate bond and its response to tensile force in hydrodynamic flow. Nature. 1995 Apr 6;374(6522):539–542. doi: 10.1038/374539a0. [DOI] [PubMed] [Google Scholar]
  3. Behrisch A., Dietrich C., Noegel A. A., Schleicher M., Sackmann E. The actin-binding protein hisactophilin binds in vitro to partially charged membranes and mediates actin coupling to membranes. Biochemistry. 1995 Nov 21;34(46):15182–15190. doi: 10.1021/bi00046a026. [DOI] [PubMed] [Google Scholar]
  4. Eichinger L., Köppel B., Noegel A. A., Schleicher M., Schliwa M., Weijer K., Witke W., Janmey P. A. Mechanical perturbation elicits a phenotypic difference between Dictyostelium wild-type cells and cytoskeletal mutants. Biophys J. 1996 Feb;70(2):1054–1060. doi: 10.1016/S0006-3495(96)79651-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Evans E. A. Detailed mechanics of membrane-membrane adhesion and separation. I. Continuum of molecular cross-bridges. Biophys J. 1985 Jul;48(1):175–183. doi: 10.1016/S0006-3495(85)83770-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Faix J., Steinmetz M., Boves H., Kammerer R. A., Lottspeich F., Mintert U., Murphy J., Stock A., Aebi U., Gerisch G. Cortexillins, major determinants of cell shape and size, are actin-bundling proteins with a parallel coiled-coil tail. Cell. 1996 Aug 23;86(4):631–642. doi: 10.1016/s0092-8674(00)80136-1. [DOI] [PubMed] [Google Scholar]
  7. Gallik S., Usami S., Jan K. M., Chien S. Shear stress-induced detachment of human polymorphonuclear leukocytes from endothelial cell monolayers. Biorheology. 1989;26(4):823–834. doi: 10.3233/bir-1989-26413. [DOI] [PubMed] [Google Scholar]
  8. Geiger B., Yehuda-Levenberg S., Bershadsky A. D. Molecular interactions in the submembrane plaque of cell-cell and cell-matrix adhesions. Acta Anat (Basel) 1995;154(1):46–62. doi: 10.1159/000147751. [DOI] [PubMed] [Google Scholar]
  9. Gingell D., Todd I. Interference reflection microscopy. A quantitative theory for image interpretation and its application to cell-substratum separation measurement. Biophys J. 1979 Jun;26(3):507–526. doi: 10.1016/S0006-3495(79)85268-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hammer D. A., Lauffenburger D. A. A dynamical model for receptor-mediated cell adhesion to surfaces. Biophys J. 1987 Sep;52(3):475–487. doi: 10.1016/S0006-3495(87)83236-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hanakam F., Gerisch G., Lotz S., Alt T., Seelig A. Binding of hisactophilin I and II to lipid membranes is controlled by a pH-dependent myristoyl-histidine switch. Biochemistry. 1996 Aug 27;35(34):11036–11044. doi: 10.1021/bi960789j. [DOI] [PubMed] [Google Scholar]
  12. Hitt A. L., Hartwig J. H., Luna E. J. Ponticulin is the major high affinity link between the plasma membrane and the cortical actin network in Dictyostelium. J Cell Biol. 1994 Sep;126(6):1433–1444. doi: 10.1083/jcb.126.6.1433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Isenberg G., Goldmann W. H. Actin-membrane coupling: a role for talin. J Muscle Res Cell Motil. 1992 Dec;13(6):587–589. doi: 10.1007/BF01738248. [DOI] [PubMed] [Google Scholar]
  15. Janmey P. A., Hvidt S., Lamb J., Stossel T. P. Resemblance of actin-binding protein/actin gels to covalently crosslinked networks. Nature. 1990 May 3;345(6270):89–92. doi: 10.1038/345089a0. [DOI] [PubMed] [Google Scholar]
  16. Jockusch B. M., Bubeck P., Giehl K., Kroemker M., Moschner J., Rothkegel M., Rüdiger M., Schlüter K., Stanke G., Winkler J. The molecular architecture of focal adhesions. Annu Rev Cell Dev Biol. 1995;11:379–416. doi: 10.1146/annurev.cb.11.110195.002115. [DOI] [PubMed] [Google Scholar]
  17. Kaufmann S., Käs J., Goldmann W. H., Sackmann E., Isenberg G. Talin anchors and nucleates actin filaments at lipid membranes. A direct demonstration. FEBS Lett. 1992 Dec 14;314(2):203–205. doi: 10.1016/0014-5793(92)80975-m. [DOI] [PubMed] [Google Scholar]
  18. Niewöhner J., Weber I., Maniak M., Müller-Taubenberger A., Gerisch G. Talin-null cells of Dictyostelium are strongly defective in adhesion to particle and substrate surfaces and slightly impaired in cytokinesis. J Cell Biol. 1997 Jul 28;138(2):349–361. doi: 10.1083/jcb.138.2.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nuckolls G. H., Romer L. H., Burridge K. Microinjection of antibodies against talin inhibits the spreading and migration of fibroblasts. J Cell Sci. 1992 Aug;102(Pt 4):753–762. doi: 10.1242/jcs.102.4.753. [DOI] [PubMed] [Google Scholar]
  20. Rädler JO, Feder TJ, Strey HH, Sackmann E. Fluctuation analysis of tension-controlled undulation forces between giant vesicles and solid substrates. Phys Rev E Stat Phys Plasmas Fluids Relat Interdiscip Topics. 1995 May;51(5):4526–4536. doi: 10.1103/physreve.51.4526. [DOI] [PubMed] [Google Scholar]
  21. Sato M., Schwarz W. H., Pollard T. D. Dependence of the mechanical properties of actin/alpha-actinin gels on deformation rate. 1987 Feb 26-Mar 4Nature. 325(6107):828–830. doi: 10.1038/325828a0. [DOI] [PubMed] [Google Scholar]
  22. Scheel J., Ziegelbauer K., Kupke T., Humbel B. M., Noegel A. A., Gerisch G., Schleicher M. Hisactophilin, a histidine-rich actin-binding protein from Dictyostelium discoideum. J Biol Chem. 1989 Feb 15;264(5):2832–2839. [PubMed] [Google Scholar]
  23. Schindl M., Wallraff E., Deubzer B., Witke W., Gerisch G., Sackmann E. Cell-substrate interactions and locomotion of Dictyostelium wild-type and mutants defective in three cytoskeletal proteins: a study using quantitative reflection interference contrast microscopy. Biophys J. 1995 Mar;68(3):1177–1190. doi: 10.1016/S0006-3495(95)80294-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Seifert U, Lipowsky R. Adhesion of vesicles. Phys Rev A. 1990 Oct 15;42(8):4768–4771. doi: 10.1103/physreva.42.4768. [DOI] [PubMed] [Google Scholar]
  25. Shutt D. C., Wessels D., Wagenknecht K., Chandrasekhar A., Hitt A. L., Luna E. J., Soll D. R. Ponticulin plays a role in the positional stabilization of pseudopods. J Cell Biol. 1995 Dec;131(6 Pt 1):1495–1506. doi: 10.1083/jcb.131.6.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tissot O., Pierres A., Foa C., Delaage M., Bongrand P. Motion of cells sedimenting on a solid surface in a laminar shear flow. Biophys J. 1992 Jan;61(1):204–215. doi: 10.1016/S0006-3495(92)81827-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Verschueren H. Interference reflection microscopy in cell biology: methodology and applications. J Cell Sci. 1985 Apr;75:279–301. doi: 10.1242/jcs.75.1.279. [DOI] [PubMed] [Google Scholar]
  28. Weber I., Wallraff E., Albrecht R., Gerisch G. Motility and substratum adhesion of Dictyostelium wild-type and cytoskeletal mutant cells: a study by RICM/bright-field double-view image analysis. J Cell Sci. 1995 Apr;108(Pt 4):1519–1530. doi: 10.1242/jcs.108.4.1519. [DOI] [PubMed] [Google Scholar]
  29. Weeks G., Herring F. G. The lipid composition and membrane fluidity of Dictyostelium discoideum plasma membranes at various stages during differentiation. J Lipid Res. 1980 Aug;21(6):681–686. [PubMed] [Google Scholar]
  30. Xia Z., Goldsmith H. L., van de Ven T. G. Flow-induced detachment of red blood cells adhering to surfaces by specific antigen-antibody bonds. Biophys J. 1994 Apr;66(4):1222–1230. doi: 10.1016/S0006-3495(94)80906-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Xia Z., Goldsmith H. L., van de Ven T. G. Kinetics of specific and nonspecific adhesion of red blood cells on glass. Biophys J. 1993 Sep;65(3):1073–1083. doi: 10.1016/S0006-3495(93)81178-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Xiao Y., Truskey G. A. Effect of receptor-ligand affinity on the strength of endothelial cell adhesion. Biophys J. 1996 Nov;71(5):2869–2884. doi: 10.1016/S0006-3495(96)79484-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zhao Y., Chien S., Skalak R. A stochastic model of leukocyte rolling. Biophys J. 1995 Oct;69(4):1309–1320. doi: 10.1016/S0006-3495(95)79998-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zilker A, Ziegler M, Sackmann E. Spectral analysis of erythrocyte flickering in the 0.3-4- microm-1 regime by microinterferometry combined with fast image processing. Phys Rev A. 1992 Dec 15;46(12):7998–8001. doi: 10.1103/physreva.46.7998. [DOI] [PubMed] [Google Scholar]
  35. van Kooten T. G., Schakenraad J. M., Van der Mei H. C., Busscher H. J. Development and use of a parallel-plate flow chamber for studying cellular adhesion to solid surfaces. J Biomed Mater Res. 1992 Jun;26(6):725–738. doi: 10.1002/jbm.820260604. [DOI] [PubMed] [Google Scholar]

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

RESOURCES