Complementary proteomics strategies capture an ataxin-1 interactome in Neuro-2a cells
Sunyuan Zhang, Nicholas A Williamson, Marie A Bogoyevitch
SCIENTIFIC DATA | NATURE PUBLISHING GROUP | Published : 2018
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. ..View full abstract
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Awarded by Australian National Health and Medical Research Council (NHMRC)
This work was funded by a grant (1121907) to M.A.B from the Australian National Health and Medical Research Council (NHMRC). S.Z. also acknowledges the scholarship support by the University of Melbourne (Melbourne International Research Scholarship). The mycBioID vector (pcDNA3.1 mycBioID) was a gift from K. Roux (addgene plasmid #35700). We acknowledge that all mass spectrometry sample preparation and analysis was performed within the Bio21 Institute Mass Spectrometry and Proteomics Facility at the University of Melbourne.