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  • 标题:Complementary proteomics strategies capture an ataxin-1 interactome in Neuro-2a cells
  • 本地全文:下载
  • 作者:Sunyuan Zhang ; Nicholas A. Williamson ; Marie A. Bogoyevitch
  • 期刊名称:Scientific Data
  • 电子版ISSN:2052-4463
  • 出版年度:2018
  • 卷号:5
  • DOI:10.1038/sdata.2018.262
  • 语种:English
  • 出版社:Nature Publishing Group
  • 摘要:Ataxin-1 mutation, arising from a polyglutamine (polyQ) tract expansion, is the underlying genetic cause of the late-onset neurodegenerative disease Spinocerebellar ataxia type 1 (SCA1). To identify protein partners of polyQ-ataxin-1 in neuronal cells under control or stress conditions, here we report our complementary proteomics strategies of proximity-dependent biotin identification (BioID) and affinity purification (via GFP-Trap pulldown) in Neuro-2a cells expressing epitope-tagged forms of ataxin-1[85Q]. These approaches allowed our enrichment of proximal proteins and interacting partners, respectively, with the subsequent protein identification performed by liquid chromatography-MS/MS. Background proteins, not dependent on the presence of the polyQ-ataxin-1 protein, were additionally defined by their endogenous biotinylation (for the BioID protocol) or by their non-specific interaction with GFP only (in the GFP-Trap protocol). All datasets were generated from biological replicates. Following the removal of the identified background proteins from the acquired protein lists, our experimental design has captured a comprehensive polyQ-ataxin-1 proximal and direct protein partners under normal and stress conditions. Data are available via ProteomeXchange, with identifier PXD010352.
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