Force induced formation and propagation of adhesion nanodomains in living fungal cells英文文献.pdfVIP

Force-induced formation and propagation of adhesion nanodomains in living fungal cells a b b,1 a,1 David Alsteens , Melissa C. Garcia , Peter N. Lipke , and Yves F. Dufrêne aInstitute of Condensed Matter and Nanosciences, Université Catholique de Louvain, B-1348 Louvain-la-Neuve, Belgium; and bDepartment of Biology, Brooklyn College of City University of New York, Brooklyn, NY 11210 Edited by Randy Schekman, University of California, Berkeley, CA, and approved October 13, 2010 (received for review September 15, 2010) Understanding how cell adhesion proteins form adhesion domains ordered adhesion domains. Whether single-molecule techniques is a key challenge in cell biology. Here, we use single-molecule can demonstrate the formation of Als5p adhesion domains in atomic force microscopy (AFM) to demonstrate the force-induced a live cell is the question that we address here. formation and propagation of adhesion nanodomains in living Single-molecule atomic force microscopy (AFM) is a powerful fungal cells, focusing on the covalently anchored cell-wall protein tool for studying how proteins respond to mechanical forces (17– Als5p from Candida albicans. We show that pulling on single adhe- 22). Stretching modular proteins such as titin (23) and tenascin sins with AFM tips terminated with specific antibodies triggers the (24) yields characteristic force signatures that reflect the force- formation of adhesion domains of 100–500 nm and that the force- induced unfolding of secondary structures (α-helices, β-sheets). induced nanodomains propagate over the entire cell surface. Con- AFM imaging has also visualized force-induced conformational trol experiments (wit