Fig. 3.

Mutational analysis. (A) Pulldown assay of SmgGDS-558 and SmgGDS-607. Pulldown results in which His-RhoA was used as bait are shown. Nonfarnesylated RhoA was used for this assay. Different mobility of RhoA observed on the SDS-PAGE was in the nature of each mutant. (B) GEF assay of SmgGDS-558 and SmgGDS-607. Guanine nucleotide dissociation was measured by monitoring fluorescence from BODYPY-GDP bound farnesylated or nonfarnesylated RhoA. The same assay was performed three times. The relative GEF activity and its SE are shown in bar chart.