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A molecular ruler regulates cytoskeletal remodelling by the Rho kinases.

Nat Commun.. 2015-12; 
Truebestein L, Elsner DJ, Fuchs E, Leonard TA.
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...SUMO1 (33–2,400, Invitrogen, 1:1,000), SUMO2/3 (M114-3, MBL,1:500), GST (A00865, GenScript, for WB analysis, 1:5,000) ...

摘要

The Rho-associated coiled-coil kinases (ROCK) are essential regulators of the actin cytoskeleton; however, the structure of a full-length ROCK is unknown and the mechanisms by which its kinase activity is controlled are not well understood. Here we determine the low-resolution structure of human ROCK2 using electron microscopy, revealing it to be a constitutive dimer, 120 nm in length, with a long coiled-coil tether linking the kinase and membrane-binding domains. We find, in contrast to previous reports, that ROCK2 activity does not appear to be directly regulated by binding to membranes, RhoA, or by phosphorylation. Instead, we show that changing the length of the tether modulates ROCK2 function in cells, s... More

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