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Single‐molecule detection of phosphorylation‐induced plasticity changes during ezrin activation
Author(s) -
Liu Dan,
Ge Ling,
Wang Fengsong,
Takahashi Hirohide,
Wang Dongmei,
Guo Zhen,
Yoshimura Shige H.,
Ward Tarsha,
Ding Xia,
Takeyasu Kunio,
Yao Xuebiao
Publication year - 2007
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2007.06.071
Subject(s) - ezrin , moesin , phosphorylation , radixin , microbiology and biotechnology , chemistry , biophysics , cytoskeleton , biology , biochemistry , cell
Ezrin–radixin–moesin protein family provides a regulated link between the cortical actin cytoskeleton and the plasma membrane. Phosphorylation of ezrin has been functionally linked to membrane dynamics and plasticity. Our recent study demonstrated that phosphorylation of the conserved T567 residue of ezrin alters the physiology of gastric parietal cells. However, the molecular mechanism of phosphorylation‐induced ezrin activation has remained elusive. Here we use atomic force microscopy (AFM) to probe phosphorylation‐mediated activation of ezrin in single molecules. The phospho‐mimicking and non‐phosphorylatable mutant ezrin proteins were generated and purified to homogeneity. Comparative analyses of two ezrin mutants by AFM demonstrate the unfolding of the N‐ and C‐terminal domains upon the phospho‐activation. To measure the physical force underlying the inter‐domain contact during mechanical unfolding, we probed the defined region of ezrin using the N‐terminal ezrin coated onto the AFM tip. Comparative force measurements indicate that T567 phosphorylation‐induced unfolding of ezrin favors the inter‐molecular association. Taken together, these results provide molecular illustration of phosphorylation elicited functional activation of ERM proteins and indicate that stimulus‐induced protein conformational change can be used as a signaling mechanism orchestrating cellular dynamics.