PINK1-Interacting Proteins: Proteomic Analysis of Overexpressed PINK1

Recent publications suggest that the Parkinson's disease- (PD-) related PINK1/Parkin pathway promotes elimination of dysfunctional mitochondria by autophagy. We used tandem affinity purification (TAP), SDS-PAGE, and mass spectrometry as a first step towards identification of possible substrates...

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Main Authors: Aleksandar Rakovic, Anne Grünewald, Lisa Voges, Sarah Hofmann, Slobodanka Orolicki, Katja Lohmann, Christine Klein
Format: Article
Language:English
Published: Wiley 2011-01-01
Series:Parkinson's Disease
Online Access:http://dx.doi.org/10.4061/2011/153979
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author Aleksandar Rakovic
Anne Grünewald
Lisa Voges
Sarah Hofmann
Slobodanka Orolicki
Katja Lohmann
Christine Klein
author_facet Aleksandar Rakovic
Anne Grünewald
Lisa Voges
Sarah Hofmann
Slobodanka Orolicki
Katja Lohmann
Christine Klein
author_sort Aleksandar Rakovic
collection DOAJ
description Recent publications suggest that the Parkinson's disease- (PD-) related PINK1/Parkin pathway promotes elimination of dysfunctional mitochondria by autophagy. We used tandem affinity purification (TAP), SDS-PAGE, and mass spectrometry as a first step towards identification of possible substrates for PINK1. The cellular abundance of selected identified interactors was investigated by Western blotting. Furthermore, one candidate gene was sequenced in 46 patients with atypical PD. In addition to two known binding partners (HSP90, CDC37), 12 proteins were identified using the TAP assay; four of which are mitochondrially localized (GRP75, HSP60, LRPPRC, and TUFM). Western blot analysis showed no differences in cellular abundance of these proteins comparing PINK1 mutant and control fibroblasts. When sequencing LRPPRC, four exonic synonymous changes and 20 polymorphisms in noncoding regions were detected. Our study provides a list of putative PINK1 binding partners, confirming previously described interactions, but also introducing novel mitochondrial proteins as potential components of the PINK1/Parkin mitophagy pathway.
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series Parkinson's Disease
spelling doaj-art-36a0a68540cb41d990deb9d7704cc1de2025-08-20T02:07:45ZengWileyParkinson's Disease2042-00802011-01-01201110.4061/2011/153979153979PINK1-Interacting Proteins: Proteomic Analysis of Overexpressed PINK1Aleksandar Rakovic0Anne Grünewald1Lisa Voges2Sarah Hofmann3Slobodanka Orolicki4Katja Lohmann5Christine Klein6Section of Clinical and Molecular Neurogenetics, Department of Neurology, University of Lübeck, Maria-Goeppert-Straße 1, 23562 Lübeck, GermanySection of Clinical and Molecular Neurogenetics, Department of Neurology, University of Lübeck, Maria-Goeppert-Straße 1, 23562 Lübeck, GermanySection of Clinical and Molecular Neurogenetics, Department of Neurology, University of Lübeck, Maria-Goeppert-Straße 1, 23562 Lübeck, GermanySection of Clinical and Molecular Neurogenetics, Department of Neurology, University of Lübeck, Maria-Goeppert-Straße 1, 23562 Lübeck, GermanySection of Clinical and Molecular Neurogenetics, Department of Neurology, University of Lübeck, Maria-Goeppert-Straße 1, 23562 Lübeck, GermanySection of Clinical and Molecular Neurogenetics, Department of Neurology, University of Lübeck, Maria-Goeppert-Straße 1, 23562 Lübeck, GermanySection of Clinical and Molecular Neurogenetics, Department of Neurology, University of Lübeck, Maria-Goeppert-Straße 1, 23562 Lübeck, GermanyRecent publications suggest that the Parkinson's disease- (PD-) related PINK1/Parkin pathway promotes elimination of dysfunctional mitochondria by autophagy. We used tandem affinity purification (TAP), SDS-PAGE, and mass spectrometry as a first step towards identification of possible substrates for PINK1. The cellular abundance of selected identified interactors was investigated by Western blotting. Furthermore, one candidate gene was sequenced in 46 patients with atypical PD. In addition to two known binding partners (HSP90, CDC37), 12 proteins were identified using the TAP assay; four of which are mitochondrially localized (GRP75, HSP60, LRPPRC, and TUFM). Western blot analysis showed no differences in cellular abundance of these proteins comparing PINK1 mutant and control fibroblasts. When sequencing LRPPRC, four exonic synonymous changes and 20 polymorphisms in noncoding regions were detected. Our study provides a list of putative PINK1 binding partners, confirming previously described interactions, but also introducing novel mitochondrial proteins as potential components of the PINK1/Parkin mitophagy pathway.http://dx.doi.org/10.4061/2011/153979
spellingShingle Aleksandar Rakovic
Anne Grünewald
Lisa Voges
Sarah Hofmann
Slobodanka Orolicki
Katja Lohmann
Christine Klein
PINK1-Interacting Proteins: Proteomic Analysis of Overexpressed PINK1
Parkinson's Disease
title PINK1-Interacting Proteins: Proteomic Analysis of Overexpressed PINK1
title_full PINK1-Interacting Proteins: Proteomic Analysis of Overexpressed PINK1
title_fullStr PINK1-Interacting Proteins: Proteomic Analysis of Overexpressed PINK1
title_full_unstemmed PINK1-Interacting Proteins: Proteomic Analysis of Overexpressed PINK1
title_short PINK1-Interacting Proteins: Proteomic Analysis of Overexpressed PINK1
title_sort pink1 interacting proteins proteomic analysis of overexpressed pink1
url http://dx.doi.org/10.4061/2011/153979
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