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An invisible ubiquitin conformation is required for efficient phosphorylation by PINK1

DOI: 10.15252/embj.201797876 DOI Help

Authors: Christina Gladkova (Medical Research Council Laboratory of Molecular Biology) , Alexander F. Schubert (Medical Research Council Laboratory of Molecular Biology) , James Wagstaff (Medical Research Council Laboratory of Molecular Biology) , Jonathan N. Pruneda (Medical Research Council Laboratory of Molecular Biology) , Stefan M. V. Freund (Medical Research Council Laboratory of Molecular Biology) , David Komander (Medical Research Council Laboratory of Molecular Biology)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: The Embo Journal

State: Published (Approved)
Published: November 2017
Diamond Proposal Number(s): 15916

Abstract: The Ser/Thr protein kinase PINK1 phosphorylates the well‐folded, globular protein ubiquitin (Ub) at a relatively protected site, Ser65. We previously showed that Ser65 phosphorylation results in a conformational change in which Ub adopts a dynamic equilibrium between the known, common Ub conformation and a distinct, second conformation wherein the last β‐strand is retracted to extend the Ser65 loop and shorten the C‐terminal tail. We show using chemical exchange saturation transfer (CEST) nuclear magnetic resonance experiments that a similar, C‐terminally retracted (Ub‐CR) conformation also exists at low population in wild‐type Ub. Point mutations in the moving β5 and neighbouring β‐strands shift the Ub/Ub‐CR equilibrium. This enabled functional studies of the two states, and we show that while the Ub‐CR conformation is defective for conjugation, it demonstrates improved binding to PINK1 through its extended Ser65 loop, and is a superior PINK1 substrate. Together our data suggest that PINK1 utilises a lowly populated yet more suitable Ub‐CR conformation of Ub for efficient phosphorylation. Our findings could be relevant for many kinases that phosphorylate residues in folded protein domains.

Journal Keywords: nuclear magnetic resonance; Parkin; Parkinson's disease; PINK1; ubiquitin phosphorylation

Diamond Keywords: Parkinson's Disease

Subject Areas: Biology and Bio-materials

Instruments: I04-Macromolecular Crystallography

Added On: 22/11/2017 12:10

Discipline Tags:

Neurodegenerative Diseases Non-Communicable Diseases Health & Wellbeing Neurology Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)