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. Author manuscript; available in PMC: 2019 Mar 16.
Published in final edited form as: J Mol Biol. 2018 Feb 1;430(6):751–758. doi: 10.1016/j.jmb.2018.01.017

Figure 3. p27 interferes with GDP exchange from RhoA induced by p115.

Figure 3

Nucleotide exchange from 1 μM MantGDP-loaded RhoA (RhoA*) in the presence of 100 μM unlabeled GDP and increasing concentrations of p27 was initiated by rapid addition of p115 to a final concentration of 1 μM. The exchange of MantGDP for unlabeled GDP was monitored by the change in fluorescence intensity over time. Panels a, b and c show titrations with p27, p27-C and p2750–105, respectively. The maximum p27 concentration tested in (a) and (b) was 0.5 mM. The potency of p2750–105 is slightly lower than that of full-length p27. The maximum concentration in (c) is 1.5 mM. The lower concentrations of p27 are 2-fold dilutions from these maxima. GDP exchange assays in the presence of p27 are depicted as circles with colors changing from red to blue to represent the serial 2-fold dilutions from the maximum concentration shown in red. The squares and triangles depicted in black are exchange assays conducted in the absence of p27 with and without p115, respectively. Error bars represent standard deviations from triplicate measurements.