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Fig. 5. Coronin 1B protects filaments from cofilin-induced depolymerization and binds antagonistically with cofilin to F-actin. (A) Time course of actin polymer concentration assembled from rabbit smooth muscle ATP G-actin, 30% pyrene labelled, after dilution from 5 µM to 0.1 µM, either alone, with phalloidin or in the presence of coronin 1B and cofilin. (B) Same experimental conditions as in A with phalloidin, coronin 1B and cofilin, or in the presence cofilin alone. (C) Representative Coomassie-Blue-stained gels showing the F-actin binding ability of coronin 1B in the presence of 2 µM cofilin, with or without pre-incubating actin filaments with 5 µM phalloidin. (D) Equilibrium binding of cofilin to actin filaments (assembled from 2 µM ATP G-actin and stored at 4°C overnight) in the presence of 1 µM coronin 1B. (E) Equilibrium binding of coronin 1B to 2 µM actin filaments in the presence of 1 µM cofilin. (F) Representative Coomassie-Blue-stained gel showing the high-speed actin co-sedimentation assay using increasing concentrations of the coronin 1B CC mutant with 2 µM actin and 1 µM cofilin.

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