Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

An ‘invisible’ ubiquitin conformation is required for efficient phosphorylation by PINK1

Christina Gladkova, Alexander F. Schubert, Jane L. Wagstaff, Jonathan N. Pruneda, Stefan M.V. Freund, David Komander
doi: https://doi.org/10.1101/189027
Christina Gladkova
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alexander F. Schubert
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jane L. Wagstaff
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jonathan N. Pruneda
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Stefan M.V. Freund
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
David Komander
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: dk@mrc-lmb.cam.ac.uk
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

ABSTRACT

The Ser/Thr protein kinase PINK1 phosphorylates the well-folded, globular protein ubiquitin (Ub) at a relatively protected site, Ser65. We had previously shown 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 in which the last β-strand is retracted to extend the Ser65 loop and shorten the C-terminal tail. We here show using Chemical Exchange Saturation Transfer (CEST) NMR experiments, that a similar, C-terminally retracted (Ub-CR) conformation exists in wild-type Ub. Ub point mutations in the moving β5-strand and in neighbouring strands shift the Ub/Ub-CR equilibrium. This enabled functional studies of the two states, and we show that the Ub-CR conformation binds to the PINK1 kinase domain 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 proteins or domains.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
Back to top
PreviousNext
Posted September 21, 2017.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
An ‘invisible’ ubiquitin conformation is required for efficient phosphorylation by PINK1
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
An ‘invisible’ ubiquitin conformation is required for efficient phosphorylation by PINK1
Christina Gladkova, Alexander F. Schubert, Jane L. Wagstaff, Jonathan N. Pruneda, Stefan M.V. Freund, David Komander
bioRxiv 189027; doi: https://doi.org/10.1101/189027
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
An ‘invisible’ ubiquitin conformation is required for efficient phosphorylation by PINK1
Christina Gladkova, Alexander F. Schubert, Jane L. Wagstaff, Jonathan N. Pruneda, Stefan M.V. Freund, David Komander
bioRxiv 189027; doi: https://doi.org/10.1101/189027

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Molecular Biology
Subject Areas
All Articles
  • Animal Behavior and Cognition (4095)
  • Biochemistry (8784)
  • Bioengineering (6490)
  • Bioinformatics (23380)
  • Biophysics (11765)
  • Cancer Biology (9166)
  • Cell Biology (13281)
  • Clinical Trials (138)
  • Developmental Biology (7421)
  • Ecology (11381)
  • Epidemiology (2066)
  • Evolutionary Biology (15111)
  • Genetics (10408)
  • Genomics (14017)
  • Immunology (9136)
  • Microbiology (22088)
  • Molecular Biology (8792)
  • Neuroscience (47421)
  • Paleontology (350)
  • Pathology (1422)
  • Pharmacology and Toxicology (2483)
  • Physiology (3710)
  • Plant Biology (8060)
  • Scientific Communication and Education (1433)
  • Synthetic Biology (2213)
  • Systems Biology (6020)
  • Zoology (1251)