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Determination of the F-actin-binding affinity of recombinant Dictyostelium EF1α domains I and III. Each recombinant protein (2 μM) was incubated with various amounts of F-actin at pH 6.6, and the amount of GST-fusion protein bound to F-actin was analyzed using actin cosedimentation assays. These data were graphed and curve fit as described in MATERIALS AND METHODS. The predicted Kds are ∼6 μM (±3 μM) and ∼0.15 μM (±0.04 μM) for domains I and domain III, respectively.
