Probes of ubiquitin E3 ligases enable systematic dissection of parkin activation

Kuan Chuan Pao, Mathew Stanley, Cong Han, Yu Chiang Lai, Paul Murphy, Kristin Balk, Nicola T. Wood, Olga Corti, Jean Christophe Corvol, Miratul M K Muqit, Satpal Virdee (Lead / Corresponding author)

Research output: Contribution to journalArticle

52 Citations (Scopus)

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 languageEnglish
Pages (from-to)324-331
Number of pages8
JournalNature Chemical Biology
Volume12
Issue number5
Early online date7 Mar 2016
DOIs
Publication statusPublished - May 2016

Fingerprint Dive into the research topics of 'Probes of ubiquitin E3 ligases enable systematic dissection of parkin activation'. Together they form a unique fingerprint.

  • Projects

    Student Theses

    Advancing understanding of endogenous PINK1-Parkin signalling linked to Parkinson’s disease

    Author: Balk, K. R., 2019

    Supervisor: Muqit, M. (Supervisor) & Alessi, D. (Supervisor)

    Student thesis: Doctoral ThesisDoctor of Philosophy

    Design and Synthesis of an E3 ligase Activity-Based Probe and its Application for the Discovery of a New Class of E3 Ligase

    Author: Pao, K., 2018

    Supervisor: Virdee, S. (Supervisor)

    Student thesis: Doctoral ThesisDoctor of Philosophy

    File

    Profiles

    No photo of Satpal Virdee

    Virdee, Satpal

    Person: Academic

    Cite this