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EMBO J. 2015 Oct 14;34(20):2492-505. doi: 10.15252/embj.201592237. Epub 2015 Aug 7.

A Ubl/ubiquitin switch in the activation of Parkin.

Author information

1
Groupe de recherché axé sur la structure des protéines and Department of Biochemistry, McGill University, Montréal, QC, Canada.
2
Department of Pharmacology and Therapeutics, McGill University, Montréal, QC, Canada.
3
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.
4
Quebec/Eastern Canada High Field NMR Facility (QANUC), Montréal, QC, Canada.
5
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.
6
Department of Pharmacology and Therapeutics, McGill University, Montréal, QC, Canada jeanfrancois.trempe@mcgill.ca kalle.gehring@mcgill.ca.
7
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.

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.

KEYWORDS:

Parkinson's disease; mitochondria; mitophagy; phosphorylation; ubiquitination

PMID:
26254305
PMCID:
PMC4609182
DOI:
10.15252/embj.201592237
[Indexed for MEDLINE]
Free PMC Article

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