Fig. 3.
AdoCbl exhibits a mixed-mode of inhibition. a Michaelis-Menten kinetics curves of full-length Invitrogen flag-tagged LRRK2-WT and c LRRK2-G2019S as a function of AdoCbl. Relative velocity represents the value of pS1292/Total LRRK2 after 20 min of reaction time at 30 °C (during the linear reaction rate), as detected by western blot and quantified by densitometry. b Lineweaver-Burk plots of LRRK2-WT and d LRRK2-G2019S kinetics data. e Microscale thermophoretic analysis of the interaction between AdoCbl and strep-tagged LRRK2 in the presence of increasing concentrations of AMP-PNP. Fluorescently-labeled strep-tagged LRRK2 was pre-incubated with AMP-PNP before binding between LRRK2 and AdoCbl was measured by MST. f Microscale thermophoretic analysis of the interaction between AMP-PNP and LRRK2, showing a KD of 0.9 μM. Data points represent the mean (±s.d.) of three biological replicates