Probes of ubiquitin E3 ligases enable systematic dissection of parkin activation
| Autor: | Satpal Virdee, Paul Murphy, Kristin Balk, Cong Han, Mathew Stanley, Yu-Chiang Lai, Kuan Chuan Pao, Miratul M. K. Muqit, Olga Corti, Jean-Christophe Corvol, Nicola T. Wood |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Cell signaling Ubiquitin-Protein Ligases Mutant medicine.disease_cause 01 natural sciences Parkin Gene Expression Regulation Enzymologic Article 03 medical and health sciences Ubiquitin medicine Humans Molecular Biology Mutation biology 010405 organic chemistry Cell Biology Fibroblasts 0104 chemical sciences 3. Good health Ubiquitin ligase Cell biology nervous system diseases 030104 developmental biology biology.protein Phosphorylation HeLa Cells |
| Zdroj: | Nature chemical biology |
| ISSN: | 1552-4469 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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