Force-induced formation and propagation of adhesion nanodomains in living fungal cells

作者: D. Alsteens , M. C. Garcia , P. N. Lipke , Y. F. Dufrene

DOI: 10.1073/PNAS.1013893107

关键词:

摘要: Understanding how cell adhesion proteins form domains is a key challenge in biology. Here, we use single-molecule atomic force microscopy (AFM) to demonstrate the force-induced formation and propagation of nanodomains living fungal cells, focusing on covalently anchored cell-wall protein Als5p from Candida albicans. We show that pulling single adhesins with AFM tips terminated specific antibodies triggers 100-500 nm propagate over entire surface. Control experiments (with cells lacking Als5p, single-site mutation protein, bare tips, modified irrelevant antibodies) result redistribution triggered by conformational changes initially probed proteins, rather than nonspecific perturbations. remodeling independent cellular metabolic activity because heat-killed same behavior as live cells. Using fluorescence microscopy, also find are formed within ∼30 min migrate at speed ∼20 nm·min(-1), indicating domain slow, time-dependent processes. These results mechanical stimuli can trigger suggest clustering may be mechanism for activating adhesion.

参考文章(30)
Marlyn Gonzalez, Piet W.J. de Groot, Frans M. Klis, Peter N. Lipke, Glycoconjugate structure and function in fungal cell walls Microbial glycobiology: structures, relevance and applications. pp. 169- 183 ,(2009) , 10.1016/B978-0-12-374546-0.00010-9
Andres F. Oberhauser, Piotr E. Marszalek, Harold P. Erickson, Julio M. Fernandez, The molecular elasticity of the extracellular matrix protein tenascin Nature. ,vol. 393, pp. 181- 185 ,(1998) , 10.1038/30270
Viola Vogel, Michael Sheetz, Local force and geometry sensing regulate cell functions Nature Reviews Molecular Cell Biology. ,vol. 7, pp. 265- 275 ,(2006) , 10.1038/NRM1890
Scott D. Gray-Owen, Richard S. Blumberg, CEACAM1: contact-dependent control of immunity. Nature Reviews Immunology. ,vol. 6, pp. 433- 446 ,(2006) , 10.1038/NRI1864
A.-S. Smith, K. Sengupta, S. Goennenwein, U. Seifert, E. Sackmann, Force-induced growth of adhesion domains is controlled by receptor mobility Proceedings of the National Academy of Sciences of the United States of America. ,vol. 105, pp. 6906- 6911 ,(2008) , 10.1073/PNAS.0801706105
M Takeichi, Cadherin cell adhesion receptors as a morphogenetic regulator Science. ,vol. 251, pp. 1451- 1455 ,(1991) , 10.1126/SCIENCE.2006419
Benjamin Geiger, Joachim P. Spatz, Alexander D. Bershadsky, Environmental sensing through focal adhesions Nature Reviews Molecular Cell Biology. ,vol. 10, pp. 21- 33 ,(2009) , 10.1038/NRM2593
Kevin J. Verstrepen, Todd B. Reynolds, Gerald R. Fink, ORIGINS OF VARIATION IN THE FUNGAL CELL SURFACE Nature Reviews Microbiology. ,vol. 2, pp. 533- 540 ,(2004) , 10.1038/NRMICRO927
Daniel J. Müller, Georg Büldt, Andreas Engel, Force-induced conformational change of bacteriorhodopsin Journal of Molecular Biology. ,vol. 249, pp. 239- 243 ,(1995) , 10.1006/JMBI.1995.0292
David Alsteens, Vincent Dupres, Stephen A. Klotz, Nand K. Gaur, Peter N. Lipke, Yves F. Dufrêne, Unfolding individual als5p adhesion proteins on live cells. ACS Nano. ,vol. 3, pp. 1677- 1682 ,(2009) , 10.1021/NN900078P