title: A Ubl/ubiquitin switch in the activation of Parkin.
abstract: Mutations in Parkin and PINK1 cause an inherited early-onset form of Parkinson's disease. The two proteins function together in a mitochondrial quality control pathway whereby PINK1 accumulates on damaged mitochondria and activates Parkin to induce mitophagy. How PINK1 kinase activity releases the auto-inhibited ubiquitin ligase activity of Parkin remains unclear. Here, we identify a binding switch between phospho-ubiquitin (pUb) and the ubiquitin-like domain (Ubl) of Parkin as a key element. By mutagenesis and SAXS, we show that pUb binds to RING1 of Parkin at a site formed by His302 and Arg305. pUb binding promotes disengagement of the Ubl from RING1 and subsequent Parkin phosphorylation. A crystal structure of Parkin Δ86-130 at 2.54 Å resolution allowed the design of mutations that specifically release the Ubl domain from RING1. These mutations mimic pUb binding and promote Parkin phosphorylation. Measurements of the E2 ubiquitin-conjugating enzyme UbcH7 binding to Parkin and Parkin E3 ligase activity suggest that Parkin phosphorylation regulates E3 ligase activity downstream of pUb binding.
authors.affiliation: Groupe de recherché axé sur la structure des protéines and Department of Biochemistry, McGill University, Montréal, QC, Canada.
authors.forename: Véronique
authors.initials: V
authors.lastname: Sauvé
authors.affiliation: Groupe de recherché axé sur la structure des protéines and Department of Biochemistry, McGill University, Montréal, QC, Canada.
authors.forename: Asparouh
authors.initials: A
authors.lastname: Lilov
authors.affiliation: Groupe de recherché axé sur la structure des protéines and Department of Biochemistry, McGill University, Montréal, QC, Canada.
authors.forename: Marjan
authors.initials: M
authors.lastname: Seirafi
authors.affiliation: Department of Pharmacology and Therapeutics, McGill University, Montréal, QC, Canada.
authors.forename: Marta
authors.initials: M
authors.lastname: Vranas
authors.affiliation: Groupe de recherché axé sur la structure des protéines and Department of Biochemistry, McGill University, Montréal, QC, Canada Department of Pharmacology and Therapeutics, McGill University, Montréal, QC, Canada.
authors.forename: Shafqat
authors.initials: S
authors.lastname: Rasool
authors.affiliation: Groupe de recherché axé sur la structure des protéines and Department of Biochemistry, McGill University, Montréal, QC, Canada.
authors.forename: Guennadi
authors.initials: G
authors.lastname: Kozlov
authors.affiliation: Quebec/Eastern Canada High Field NMR Facility (QANUC), Montréal, QC, Canada.
authors.forename: Tara
authors.initials: T
authors.lastname: Sprules
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.
authors.forename: Jimin
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Pharmacology and Therapeutics, McGill University, Montréal, QC, Canada jeanfrancois.trempe@mcgill.ca kalle.gehring@mcgill.ca.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Trempe
authors.affiliation: Groupe de recherché axé sur la structure des protéines and Department of Biochemistry, McGill University, Montréal, QC, Canada jeanfrancois.trempe@mcgill.ca kalle.gehring@mcgill.ca.
authors.forename: Kalle
authors.initials: K
authors.lastname: Gehring
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D004789:Enzyme Activation / Q000235:genetics*; D006801:Humans; D008928:Mitochondria / Q000378:metabolism*; D008958:Models, Molecular; D016296:Mutagenesis; D019906:Nuclear Magnetic Resonance, Biomolecular; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D053838:Scattering, Small Angle; D025801:Ubiquitin / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry* / Q000378:metabolism*; D000099202:PTEN-Induced Putative Kinase
chemical_list: D011494:Protein Kinases; D025801:Ubiquitin; D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases; D000099202:PTEN-Induced Putative Kinase; C079437:UBE2L3 protein, human; C111567:parkin protein
keywords: Parkinson's disease; mitochondria; mitophagy; phosphorylation; ubiquitination