PINK1 is activated by mitochondrial membrane potential depolarization and stimulates Parkin E3 ligase activity by phosphorylating Serine 65

By Chandana Kondapalli, Agne Kazlauskaite, Ning Zhang, Helen I. Woodroof, D. Campbell, Robert Gourlay, L. Burchell, H. Walden, Thomas J. Macartney, M. Deák and 3 more. Open Biology, 2012

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Summary Missense mutations in PTEN-induced kinase 1 (PINK1) cause autosomal-recessive inherited Parkinson's disease (PD). We have exploited our recent discovery that recombinant insect PINK1 is catalytically active to test whether PINK1 directly phosphorylates 15 proteins encoded by PD-associated genes as well as proteins reported to bind PINK1. We have discovered that insect PINK1 efficiently phosphorylates only one of these proteins, namely the E3 ligase Parkin. We have mapped the phosphorylation site to a highly conserved residue within the Ubl domain of Parkin at Ser65. We show that human PINK1 is specifically activated by mitochondrial membrane potential (Δψm) depolarization, enabling it to phosphorylate Parkin at Ser65. We further show that phosphorylation of Parkin at Ser65 leads to marked activation of its E3 ligase activity that is prevented by mutation of Ser65 or inactivation of PINK1. We provide evidence that once activated, PINK1 autophosphorylates at several residues, including Thr257, which is accompanied by an electrophoretic mobility band-shift. These results provide the first evidence that PINK1 is activated following Δψm depolarization and suggest that PINK1 directly phosphorylates and activates Parkin. Our findings indicate that monitoring phosphorylation of Parkin at Ser65 and/or PINK1 at Thr257 represent the first biomarkers for examining activity of the PINK1-Parkin signalling pathway in vivo. Our findings also suggest that small molecule activators of Parkin that mimic the effect of PINK1 phosphorylation may confer therapeutic benefit for PD.

Subfields: Cell Biology, Epidemiology, Neurology

View Paper: https://www.semanticscholar.org/paper/acbdb98143276b513c482aa833f06abac8f6ef49

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Citations in the corpus

Quantitative proteomics reveal a feed-forward model for mitochondrial PARKIN translocation and UB chain synthesis (2014, positive): “Phosphorylation of PARKIN and UB by PINK1Tc (Kondapalli et al., 2012) was performed as described in Supplemental Experimental Procedures.”
A new pathway for mitochondrial quality control: mitochondrial‐derived vesicles (2014, background): “…matrix processing peptidases and PARL (Jin et al, 2010; Greene et al, 2012); however, almost all of the cleaved PINK1 is then released from the import channel and degraded in the cytosol through the N-end rule proteolytic pathway (Kondapalli et al, 2012; Lazarou et al, 2012; Yamano & Youle, 2013).”
Landscape of the PARKIN-dependent ubiquitylome in response to mitochondrial depolarization (2013, background): “…UBL domain (residues 1–80), 2) K27/48R, which blocks ubiquitylation of two regulated diGLY sites in the UBL (Figure 2), 3) S65A and S65E, which block PINK1-dependent phosphorylation of the UBL domain implicated in PARKIN activation in vitro, 18 and 4) C431S, removing the active site Cys in RING2.”
Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) (2012, neutral): “Moreover, mitophagy elicited in a number of disease models does not involve mitochondrial PARK2 translocation.(145,347,785) Along these lines, recent studies implicate an essential role for TRAF2, an E3 ubiquitin ligase, as a mitophagy effector in concert with PARK2 in cardiac myocytes; whereby mito”
Deciphering the Molecular Signals of PINK1/Parkin Mitophagy. (2016, background): “PINK1 phosphorylates Parkin at S65 in the UBL domain and this was shown to stimulate Parkin's ligase activity and recruitment to mitochondria [15,16].”
The PINK1-PARKIN mitochondrial ubiquitylation pathway drives a program of TBK1 activation and recruitment of OPTN and NDP52 to promote mitophagy (2015, background): “…on the mitochondrial outer membrane (MOM), where it promotes phosphorylation of PARKIN on S65 in its N-terminal UB-like domain, as well as the conserved S65 within UB on mitochondria (Kane et al., 2014; Kazlauskaite et al., 2014; Kondapalli et al., 2012; Koyano et al., 2014; Ordureau et al., 2014).”