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Human in vitro Parkin ubiquitin E3 ligase assay v1
[dataset]
2021
protocols.io
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Mutations in PARK2 encoding Parkin are causal for early-onset Parkinson's disease. Parkin is a ubiquitin E3 ligase and is activated by the PINK1 kinase to stimulate ubiquitin-dependent removal of damaged mitochondria via autophagy (mitophagy). Previous studies have identified Miro1 as a Parkin substrate in cells and in vitro. This protocol describes a facile direct assay of Parkin E3 ligase activity using recombinant untagged Parkin and GST-Miro1 as a substrate. This assay can be employed to
doi:10.17504/protocols.io.bsfrnbm6
fatcat:bwqbfzidj5d6tdw36athvpahsa