Figure 3

Putative Ub-binding groove of TcPINK1. (A) In vitro kinase assays of TcPINK1 mutants that were expected to compromise the interaction with Ub. The Ub phosphorylation (pUb) by wild-type or mutant GST-TcPINK1 was analysed by Phos-tag SDS-PAGE with Coomassie brilliant blue staining. The colour-coded asterisks represent the magnitude of the effect of each mutation on the Ub phosphorylation: red, drastic (less than one-tenth dose of wild type); orange, mild (less than wild type); blue, little or no (nearly equal to wild type). (B,C) Mapping of potential Ub-interacting residues on the molecular surface (B) and cartoon model (C) of TcPINK1^DDEE. The potential Ub-interacting residues are shown as sticks in (C). The colour-coding is based on the magnitude of the effect of each mutation on Ub phosphorylation, except that phosphomimetic sites are coloured in purple: red, drastic (less than one-tenth dose of wild type); orange, mild (less than wild type); blue, little or no (nearly equal to wild type).