Figure 6: Parkin is not a substrate of parkin in trans.
From: A molecular explanation for the recessive nature of parkin-linked Parkinson’s disease
(a) Ubiquitination assay using IBR–RING2–T415N (IBR–R2–T415N) as a substrate. Each parkin species point mutation is assayed in the absence (−) or presence (+) of substrate. The first lane shows wild-type active IBR–RING2, the second lane shows the inactive mutant. Parkin–ubiquitin conjugates are detected by western blotting using a parkin antibody. (b) K27N–parkin is assayed for activity towards IBR-R2-T415N with increasing substrate concentration. Parkin–ubiquitin conjugates are detected by western blotting using parkin (left) and His-ubiquitin (right) antibodies. (c) Ubiquitination assay using full-length HA–parkin–T415N as a substrate. Parkin–ubiquitin conjugates are detected by western blotting using parkin and His-ubiquitin antibodies. Unmodified HA-parkin-T415N is detected using HA antibodies. The asterisk denotes a non-specific band.
