Profiling of Parkin-Binding Partners Using Tandem Affinity Purification

Parkinson's disease (PD) is a progressive neurodegenerative disorder affecting approximately 1-2% of the general population over age 60. It is characterized by a rather selective loss of dopaminergic neurons in the substantia nigra and the presence of α-synuclein-enriched Lewy body inclusions....

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Vydáno v:	PloS one Ročník 8; číslo 11; s. e78648
Hlavní autoři:	Zanon, Alessandra, Rakovic, Aleksandar, Blankenburg, Hagen, Doncheva, Nadezhda T., Schwienbacher, Christine, Serafin, Alice, Alexa, Adrian, Weichenberger, Christian X., Albrecht, Mario, Klein, Christine, Hicks, Andrew A., Pramstaller, Peter P., Domingues, Francisco S., Pichler, Irene
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	United States Public Library of Science 11.11.2013 Public Library of Science (PLoS)
Témata:	Acids Affinity Age Animals Basal ganglia Binding proteins Bioinformatics Biological activity Biology Cell cycle Cell death Cell Line, Tumor Central nervous system diseases Depolarization Dopamine Dopamine receptors Drosophila Drosophila melanogaster Drosophila Proteins - genetics Drosophila Proteins - isolation & purification Drosophila Proteins - metabolism Fusion protein Gene expression Genes Genetic aspects Health screening Humans Immunoprecipitation Informatics Insects Lewy bodies Machinery Machinery and equipment Mass spectrometry Mass spectroscopy Mitochondria Movement disorders Mutation Nerve Tissue Proteins - genetics Nerve Tissue Proteins - isolation & purification Nerve Tissue Proteins - metabolism Nervous system diseases Neurodegeneration Neurodegenerative diseases Neurons Ontology Parkin protein Parkinson disease Parkinson Disease - genetics Parkinson Disease - metabolism Parkinson's disease Protein Binding Protein folding Protein interaction Protein purification Protein-protein interactions Proteins Purification Screens Substantia nigra Synuclein Ubiquitin Ubiquitin-protein ligase Ubiquitin-Protein Ligases - genetics Ubiquitin-Protein Ligases - isolation & purification Ubiquitin-Protein Ligases - metabolism Italy Germany
ISSN:	1932-6203, 1932-6203
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Shrnutí:	Parkinson's disease (PD) is a progressive neurodegenerative disorder affecting approximately 1-2% of the general population over age 60. It is characterized by a rather selective loss of dopaminergic neurons in the substantia nigra and the presence of α-synuclein-enriched Lewy body inclusions. Mutations in the Parkin gene (PARK2) are the major cause of autosomal recessive early-onset parkinsonism. The Parkin protein is an E3 ubiquitin ligase with various cellular functions, including the induction of mitophagy upon mitochondrial depolarizaton, but the full repertoire of Parkin-binding proteins remains poorly defined. Here we employed tandem affinity purification interaction screens with subsequent mass spectrometry to profile binding partners of Parkin. Using this approach for two different cell types (HEK293T and SH-SY5Y neuronal cells), we identified a total of 203 candidate Parkin-binding proteins. For the candidate proteins and the proteins known to cause heritable forms of parkinsonism, protein-protein interaction data were derived from public databases, and the associated biological processes and pathways were analyzed and compared. Functional similarity between the candidates and the proteins involved in monogenic parkinsonism was investigated, and additional confirmatory evidence was obtained using published genetic interaction data from Drosophila melanogaster. Based on the results of the different analyses, a prioritization score was assigned to each candidate Parkin-binding protein. Two of the top ranking candidates were tested by co-immunoprecipitation, and interaction to Parkin was confirmed for one of them. New candidates for involvement in cell death processes, protein folding, the fission/fusion machinery, and the mitophagy pathway were identified, which provide a resource for further elucidating Parkin function.
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Current address: Illumina, Cambridge Ltd, Cambridge, United Kingdom Competing Interests: The authors have read the journal's policy and have the following conflicts: Peter P. Pramstaller received honoraria for serving on scientific boards and speaking from Novartis, Boehringer, GlaxoSmithKline, Lundbeck and UCB. Christine Klein is a member of the editorial board of “Neurology” and has served as editor of the “Continuum Issue Neurogenetics 2008” and as faculty at the Annual Meetings of the American Academy of Neurology since 2004. She is a consultant to Centogene and received honoraria for speaking from Boehringer Ingelheim and Orion Pharma. Dr. Klein is the recipient of a career development award from the Hermann and Lilly Schilling Foundation. She is funded by the Volkswagen Foundation, the Deutsche Forschungsgemeinschaft, the Possehl Foundation and received institutional support from the University of Luebeck for genetics research. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials. Also, the current affiliation of Adrian Alexa, one of the co-authors of the paper, to Illumina, Cambridge Ltd, UK (stated on the first page of the manuscript) does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials. Conceived and designed the experiments: CK PPP AAH IP FSD. Performed the experiments: AZ AS IP AR CS. Analyzed the data: HB NTD CXW FSD. Contributed reagents/materials/analysis tools: AA MA. Wrote the paper: IP AZ FSD.
ISSN:	1932-6203 1932-6203
DOI:	10.1371/journal.pone.0078648