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. 2015 Sep 23;30(1):336–347. doi: 10.1096/fj.15-277095

Figure 2.

Figure 2.

ROC phosphorylation enhances its GTPase activity. A) Comparison of kinetics of GTP hydrolysis by ROC and pROC. Equal concentrations (20 μM) of His6-ROC or His6-pROC were assayed for GTP hydrolysis by using the various concentrations of GTP indicated. Data were fit to nonlinear regression kcat curves, and the rate constants were determined. B) Time course for GTP-hydrolysis by His6-ROC or His6-pROC (each at 20 μM) in the presence of saturating (1 mM) GTP. GTP off rates of His6-ROC and His6-pROC proteins (each at 0.3 μM) were measured using α-[32P]GTP (C) or [3H]GDP (D) and found to be indistinguishable. E) Time course for GTP hydrolysis by GST-[Δ970] LRRK2G2019S or pGST-[Δ970] LRRK2G2019S (each at 2 μM) in the presence of saturating (1 mM) GTP. All rate constants were calculated from the average of 3 independent experiments. Data are means ± sem; significances by unpaired Student’s t test.