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Figure 2.
Structural basis of PD-associated mutations in ROC. (A) R1441
and W1434 from one monomer together with F1401 and P1406 from
the other stack on each other alternately, forming a hydrophobic
zipper at the dimer interface. The guanidinium group of R1441
also is hydrogen-bonded with the backbone carbonyl oxygen of
F1401 and the hydroxyl group of T1404 on helix α2 from the
other peptide chain. 2mF [o] − DF [c] electron density map is
shown in blue. (B) I1371 is inserted in a hydrophobic cavity,
which is constructed by residues from both monomers at the dimer
interface. I1371 is shown in stick format and colored in orange.
The surrounding residues are shown in stick format within the
semitransparent surface representation. The color scheme is the
same as that in Fig. 1. Note the side-chain methyl group of
T1404 is pointing directly to the tip of I1371, forming a
favorable van der Waals' interaction. (C) R1441C (lane 3), as a
prototypical mutation at the dimer interface, decreases
interaction with the full-length wild-type LRRK2 protein
compared with wild-type GST fusions (lane 2); no interaction was
seen with GST alone (lane 1). (D) Pull-down assays were
quantified and corrected for the amount of LRRK2 protein in the
inputs (middle blots). *, P < 0.0001; **, P < 0.01 compared with
GST alone (one-way ANOVA with Student–Newman–Kuell's post
hoc test; n = 3).
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