Projects per year
Abstract
E3 ligases represent an important class of enzymes, yet there are currently no chemical probes for profiling their activity. We develop a new class of activity-based probe by re-engineering a ubiquitin-charged E2 conjugating enzyme and demonstrate the utility of these probes by profiling the transthiolation activity of the RING-in-between-RING (RBR) E3 ligase parkin in vitro and in cellular extracts. Our study provides valuable insight into the roles, and cellular hierarchy, of distinct phosphorylation events in parkin activation. We also profile parkin mutations associated with patients with Parkinson's disease and demonstrate that they mediate their effect largely by altering transthiolation activity. Furthermore, our probes enable direct and quantitative measurement of endogenous parkin activity, revealing that endogenous parkin is activated in neuronal cell lines (≥75%) in response to mitochondrial depolarization. This new technology also holds promise as a novel biomarker of PINK1-parkin signaling, as demonstrated by its compatibility with samples derived from individuals with Parkinson's disease.
Original language | English |
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Pages (from-to) | 324-331 |
Number of pages | 8 |
Journal | Nature Chemical Biology |
Volume | 12 |
Issue number | 5 |
Early online date | 7 Mar 2016 |
DOIs | |
Publication status | Published - May 2016 |
ASJC Scopus subject areas
- Cell Biology
- Molecular Biology
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Dive into the research topics of 'Probes of ubiquitin E3 ligases enable systematic dissection of parkin activation'. Together they form a unique fingerprint.Projects
- 2 Finished
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Structural Analysis of the Parkinson's Associated Kinase PINK1
Muqit, M. (Investigator) & van Aalten, D. (Investigator)
1/11/15 → 31/10/16
Project: Research
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Investigation of the Regulation of Rab GTPases by PINK1 (PhD Studentship)
Muqit, M. (Investigator)
7/09/15 → 6/09/18
Project: Research
Student theses
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Advancing understanding of endogenous PINK1-Parkin signalling linked to Parkinson’s disease
Balk, K. R. (Author), Muqit, M. (Supervisor) & Alessi, D. (Supervisor), 2019Student thesis: Doctoral Thesis › Doctor of Philosophy
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Design and Synthesis of an E3 ligase Activity-Based Probe and its Application for the Discovery of a New Class of E3 Ligase
Pao, K.-C. (Author), Virdee, S. (Supervisor), 2018Student thesis: Doctoral Thesis › Doctor of Philosophy
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