Fig. 3

Both disease-causing mutant PINK1 protein and Ser465A mutant PINK1 protein decreased its kinase activity. a Sanger sequencing of T313 M mutant PINK1 and WT PINK1. The arrow indicates that the 938th base mutated from C to T, leading to threonine mutated to methionine. b Sanger sequencing of R492X mutant PINK1 and WT PINK1. The arrow indicates that the 1474th base mutated from C to T, leading to arginine mutated to a stop code. (c, d) Coomassie blue staining of purified GST-PINK1-T313 M and GST-PINK1-R492X fusion proteins. e Wild-type and three mutant GST-PINK1 proteins phosphorylation activity with casein as the common substrate. The upper one is the result of Coomassie Blue staining. Each lane represents GST-PINK1-WT fusion protein, GST protein, GST-PINK1-T313 M fusion protein and GST-PINK1-R492X fusion protein, respectively. The lower one was the result of autoradiography of five groups. Except GST group, four positive signals appeared in the other four groups, standing for the casein phosphorylated by GST-PINK1-WT fusion protein, GST-PINK1-T313 M fusion protein, GST-PINK1-R492X fusion protein and GST-PINK1-S465A fusion protein respectively. f Bar graph showing the quantification of phosphorylation activity of the five groups. The difference between three mutant PINK1 proteins and WT protein were statistically significant (^* P < 0.05)