Proteomic analysis of increased Parkin expression and its interactants provides evidence for a role in modulation of mitochondrial function

Davison, Eleanor J. and Pennington, Kyla and Hung, Chao-Chung and Peng, Jianhe and Rafiq, Rumana and Ostareck-Lederer, Dirk H. and Ardley, Helen C. and Banks, Rosamonde E. and Robinson, Philip A. (2009) Proteomic analysis of increased Parkin expression and its interactants provides evidence for a role in modulation of mitochondrial function. Proteomics, 9 (18). pp. 4284-4297. ISSN 1615-9853

Full content URL: http://onlinelibrary.wiley.com/doi/10.1002/pmic.20...

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Item Type:Article
Item Status:Live Archive

Abstract

Parkin is an ubiquitin-protein ligase (E3), mutations of which cause juvenile onset – autosomal recessive Parkinson's disease, and result in reduced enzymic activity. In contrast, increased levels are protective against mitochondrial dysfunction and neurodegeneration, the mechanism of which is largely unknown. In this study, 2-DE and MS proteomic techniques were utilised to investigate the effects of increased Parkin levels on protein expression in whole cell lysates using in an inducible Parkin expression system in HEK293 cells, and also to isolate potential interactants of Parkin using tandem affinity purification and MS. Nine proteins were significantly differentially expressed (±2-fold change; p<0.05) using 2-DE analysis. MS revealed the identity of these proteins to be ACAT2, HNRNPK, HSPD1, PGK1, PRDX6, VCL, VIM, TPI1, and IMPDH2. The first seven of these were reduced in expression. Western blot analysis confirmed the reduction in one of these proteins (HNRNPK), and that its levels were dependent on 26S proteasomal activity. Tandem affinity purification/MS revealed 14 potential interactants of Parkin; CKB, DBT, HSPD1, HSPA9, LRPPRC, NDUFS2, PRDX6, SLC25A5, TPI1, UCHL1, UQCRC1, VCL, YWHAZ, YWHAE. Nine of these are directly involved in mitochondrial energy metabolism and glycolysis; four were also identified in the 2-DE study (HSP60, PRDX6, TPI1, and VCL). This study provides further evidence for a role for Parkin in regulating mitochondrial activity within cells.

Keywords:2-DE, Cell biology, Mitochondria, Parkin, Parkinson's disease, Ubiquitination
Subjects:C Biological Sciences > C800 Psychology
C Biological Sciences > C850 Cognitive Psychology
Divisions:College of Social Science > School of Psychology
ID Code:11758
Deposited On:02 Sep 2013 16:15

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