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Figure 4.
Figure 4: Mutually exclusive binding of Rho and DAD to mDia[N].
a, b, Observed association rate constants for the binding of
wild-type and mutant RhoA to wild-type mDia[N] (a) or wild-type
RhoA to different mDia[N] mutants (b), measured as in Fig. 1. c,
d, Fluorescence polarization assay with 1 µM AMCA-labelled DAD
peptide, mDia[N] mutants A256D (c) and N165D (d) and RhoA as
described in Fig. 2. In c, wild-type mDia[N] was added as a
control, to show DAD binding ability. All experiments were
repeated at least twice and typical experiments are shown. e,
Schematic diagram of the partially overlapping binding sites
identified here and the proposed mechanism of release of
DAD-mediated autoinhibition by Rho, as discussed in the text.
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