Effects of LD mutations on the interaction of laforin with malin and PTG
(A) Interaction of WT laforin and mutants with GAD, GAD-PTG, and GAD-malin fusion proteins. The dotted line indicates WT level of malin interaction, and the dashed line indicates WT level of PTG interaction.
(B) Triple hybrid experiment demonstrating noncompetitive binding of malin and PTG to laforin. LexA-laforin was expressed with GAD-malin and pWS-PTG or with GAD-PTG and pWS-malin. In (A) and (B) bar graphs represent the average of triplicate reactions ±SD.
(C–G) Correlation between Tm and GAD-malin interaction (r = 0.6087, p = 0.0021), Tm and GAD-PTG interaction (r = 0.499, p = 0.0154), GAD-malin and GAD-PTG interactions (r = 0.6462, p = 0.0009), glycogen activity and GAD-malin interaction (r = 0.01087, p = 0.9607), and glycogen activity and GAD-PTG interaction (r = 0.3646, p = 0.0872). In e, inset corresponds to the boxed region. Dot color corresponds to mutation location. See Figure S3 for the full 2D correlation analysis and Table S1 for all correlation coefficients and p values.