Fig. 2

Biochemical properties of EsuRRAS2-like and HsaRRAS2 proteins. A Titration of EsuRRAS2-like and HsaRRAS2 proteins in GTPase/GAP Buffer with a molar concentration of 1 µM GTP. Successful isolation and purification of EsuRRAS2-like and HsaRRAS2 proteins confirmed by SDS-PAGE gel. Concentration of 6.25 µM (791.875 ng) is marked with rectangle. B Both the sponge and human R-RAS2 homologs exhibited intrinsic GTPase activity at a concentration of 6.25 µM. Luminescence was measured after a two-hour incubation. The control sample contained only the GTP/GAP buffer. Standard deviations are indicated as mean ± SD, n = 3. RLU (relative luminescence unit). ***p < 0.005. C RNA binding activity of sponge R-RAS2-like and human R-RAS2. Sponge and human proteins (5 µg) were preincubated with increasing concentrations of free poly(U), followed by incubation with 50% poly(U) agarose beads. The RNA binding protein DRG1 served as a positive control, while BSA served as a negative control. After incubation, proteins were analyzed by SDS-PAGE and stained with Coomassie Brilliant Blue. Abbreviations: I-input, B-beads