G2385R and I2020T Mutations Increase LRRK2 GTPase Activity

Figure 2

In vitro kinase assays with commercially available recombinant LRRK2 proteins. The assays were carried out using purified recombinant His-Rab5B protein as the substrate. (a) LRRK2 and Rab5B phosphorylations were shown by autoradiography. The quantities of the LRRK2 and Rab5B proteins used are shown in the Western blot results with the GST and Rab5B antibodies below the corresponding autoradiograms. Relative Rab5B phosphorylation was calculated and is shown below the Rab5B Western blot. (b) Relative kinase activities of autophosphorylated LRRK2 after a statistical analysis of three independent experiments. All values are relative to the WT and are expressed as the mean ± SEM. —: no added LRRK2 protein, WT: wild type, GS: G2019S, DA: D1994A, RC: R1441C, IT: I2020T, and GR: G2385R. and .

(a)

(b)