PINK1 is a mitochondrial kinase responsible for initiating the selective autophagic elimination of damaged mitochondria, a process known as mitophagy. Defects in PINK1 lead to impaired mitochondrial function and the development of early onset Parkinson’s disease (EOPD). Therefore, enhancing PINK1 activity to augment mitochondrial health may be a promising therapeutic strategy to treat EOPD.
We have recently shown that, in vitro, Pediculus humanus corporis PINK1 (PhPINK1) homo-oligomerises into a dodecamer, enabling structural determination of PINK1 by cryo-EM (Gan et al., Nature 602, 328–335, 2022). While the arrangement of PhPINK1 molecules within the dodecamer provided insights into PINK1 activation, the PhPINK1 dodecamer itself, with exposed and unobstructed ATP binding sites, may provide a straightforward method to solve ligand-bound structures of PINK1. Here, we determined nucleotide-bound structures of PhPINK1 after incubation of the dodecamer with nucleotides prior to vitrification and cryo-EM, confirming the feasibility of this approach. The structures further reveal that conformation changes in the kinase occur upon nucleotide binding. The PhPINK1 dodecamer therefore enables the rapid determination of ligand-bound structures of PINK1 and may further allow the determination of PINK1 structures in complex with potential therapeutic activators of PINK1 activity.