Structural insights into Parkin substrate lysine targeting from minimal Miro substrates

Julian L. Klosowiak, Sungjin Park, Kyle P. Smith, Michael E. French, Pamela J Focia, Douglas M Freymann, Sarah E Rice*

*Corresponding author for this work

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Hereditary Parkinson's disease is commonly caused by mutations in the protein kinase PINK1 or the E3 ubiquitin ligase Parkin, which function together to eliminate damaged mitochondria. PINK1 phosphorylates both Parkin and ubiquitin to stimulate ubiquitination of dozens of proteins on the surface of the outer mitochondrial membrane. However, the mechanisms by which Parkin recognizes specific proteins for modification remain largely unexplored. Here, we show that the C-terminal GTPase (cGTPase) of the Parkin primary substrate human Miro is necessary and sufficient for efficient ubiquitination. We present several new X-ray crystal structures of both human Miro1 and Miro2 that reveal substrate recognition and ubiquitin transfer to be specific to particular protein domains and lysine residues. We also provide evidence that Parkin substrate recognition is functionally separate from substrate modification. Finally, we show that prioritization for modification of a specific lysine sidechain of the cGTPase (K572) within human Miro1 is dependent on both its location and chemical microenvironment. Activation of Parkin by phosphorylation or by binding of pUb is required for prioritization of K572 for modification, suggesting that Parkin activation and acquisition of substrate specificity are coupled.

Original languageEnglish (US)
Article number33019
JournalScientific Reports
Volume6
DOIs
StatePublished - Sep 8 2016

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Lysine
Ubiquitination
GTP Phosphohydrolases
Ubiquitin
Inborn Genetic Diseases
Ubiquitin-Protein Ligases
Mitochondrial Membranes
Substrate Specificity
Protein Kinases
Parkinson Disease
Mitochondria
Membrane Proteins
Phosphorylation
X-Rays
Mutation
Proteins
Protein Domains

ASJC Scopus subject areas

  • General

Cite this

Klosowiak, Julian L. ; Park, Sungjin ; Smith, Kyle P. ; French, Michael E. ; Focia, Pamela J ; Freymann, Douglas M ; Rice, Sarah E. / Structural insights into Parkin substrate lysine targeting from minimal Miro substrates. In: Scientific Reports. 2016 ; Vol. 6.
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Structural insights into Parkin substrate lysine targeting from minimal Miro substrates. / Klosowiak, Julian L.; Park, Sungjin; Smith, Kyle P.; French, Michael E.; Focia, Pamela J; Freymann, Douglas M; Rice, Sarah E.

In: Scientific Reports, Vol. 6, 33019, 08.09.2016.

Research output: Contribution to journalArticle

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