Characterization of PINK1 processing, stability, and subcellular localization

William Lin, Un Jung Kang

Research output: Contribution to journalArticle

147 Citations (Scopus)

Abstract

Mutations found in PTEN-induced putative kinase 1 (PINK1), a putative mitochondrial serine/threonine kinase of unknown function, have been linked to autosomal recessive Parkinson's disease. It is suggested that mutations can cause a loss of PINK1 kinase activity and eventually lead to mitochondrial dysfunction. In this report, we examined the subcellular localization of PINK1 and the dynamic kinetics of PINK1 processing and degradation. We also identified cytosolic chaperone heat-shock protein 90 (Hsp90) as an interacting protein of PINK1 by PINK1 co-immunoprecipitation. Immunofluorescence of PINK1 protein and mitochondrial isolation show that the precursor form of PINK1 translocates to the mitochondria and is processed into two cleaved forms of PINK1, which in turn localize more to the cytosolic than mitochondrial fraction. The cleavage does not occur and the uncleaved precursor stays associated with the mitochondria when the mitochondrial membrane potential is disrupted. Metabolic labeling analyses show that the PINK1 processing is rapid and the levels of cleaved forms are tightly regulated. Furthermore, cleaved forms of PINK1 are stabilized by Hsp90 interaction as the loss of Hsp90 activity decreases PINK1 level after mitochondrial processing. Lastly, we also find that cleaved forms of PINK1 are degraded by the proteasome, which is uncommon for mitochondrial proteins. Our findings support a dual subcellular localization, implying that PINK1 can reside in the mitochondria and the cytosol. This raises intriguing functional roles that bridge these two cellular compartments.

Original languageEnglish
Pages (from-to)464-474
Number of pages11
JournalJournal of Neurochemistry
Volume106
Issue number1
DOIs
StatePublished - 1 Jul 2008

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Processing
HSP90 Heat-Shock Proteins
Mitochondria
Mitochondrial Proteins
PTEN-induced putative kinase
Mutation
Mitochondrial Membrane Potential
Protein-Serine-Threonine Kinases
Proteasome Endopeptidase Complex
Immunoprecipitation
Cytosol
Labeling
Fluorescent Antibody Technique
Parkinson Disease
Phosphotransferases
Proteins
Membranes
Degradation
Kinetics

Keywords

  • Heat-shock protein 90
  • Mitochondria
  • PTEN-induced putative kinase 1
  • Parkinson's disease
  • Valinomycin

Cite this

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abstract = "Mutations found in PTEN-induced putative kinase 1 (PINK1), a putative mitochondrial serine/threonine kinase of unknown function, have been linked to autosomal recessive Parkinson's disease. It is suggested that mutations can cause a loss of PINK1 kinase activity and eventually lead to mitochondrial dysfunction. In this report, we examined the subcellular localization of PINK1 and the dynamic kinetics of PINK1 processing and degradation. We also identified cytosolic chaperone heat-shock protein 90 (Hsp90) as an interacting protein of PINK1 by PINK1 co-immunoprecipitation. Immunofluorescence of PINK1 protein and mitochondrial isolation show that the precursor form of PINK1 translocates to the mitochondria and is processed into two cleaved forms of PINK1, which in turn localize more to the cytosolic than mitochondrial fraction. The cleavage does not occur and the uncleaved precursor stays associated with the mitochondria when the mitochondrial membrane potential is disrupted. Metabolic labeling analyses show that the PINK1 processing is rapid and the levels of cleaved forms are tightly regulated. Furthermore, cleaved forms of PINK1 are stabilized by Hsp90 interaction as the loss of Hsp90 activity decreases PINK1 level after mitochondrial processing. Lastly, we also find that cleaved forms of PINK1 are degraded by the proteasome, which is uncommon for mitochondrial proteins. Our findings support a dual subcellular localization, implying that PINK1 can reside in the mitochondria and the cytosol. This raises intriguing functional roles that bridge these two cellular compartments.",
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Characterization of PINK1 processing, stability, and subcellular localization. / Lin, William; Kang, Un Jung.

In: Journal of Neurochemistry, Vol. 106, No. 1, 01.07.2008, p. 464-474.

Research output: Contribution to journalArticle

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