1. A Ubl/ubiquitin switch in the activation of Parkin.
- Author
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Sauvé V, Lilov A, Seirafi M, Vranas M, Rasool S, Kozlov G, Sprules T, Wang J, Trempe JF, and Gehring K
- Subjects
- Cloning, Molecular, Crystallization, Humans, Mutagenesis, Nuclear Magnetic Resonance, Biomolecular, Phosphorylation, Protein Binding, Protein Conformation, Protein Structure, Tertiary, Scattering, Small Angle, Ubiquitin metabolism, Ubiquitin-Conjugating Enzymes metabolism, Enzyme Activation genetics, Mitochondria metabolism, Models, Molecular, Protein Kinases chemistry, Protein Kinases metabolism, Ubiquitin-Protein Ligases chemistry, Ubiquitin-Protein Ligases metabolism
- 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., (© 2015 The Authors. Published under the terms of the CC BY 4.0 license.)
- Published
- 2015
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