Tuning Hsp104 specificity to selectively detoxify α-synuclein

Korrie L. Mack, Hanna Kim, Edward M. Barbieri, Jia Bei Lin, Sylvanne Braganza, Meredith E. Jackrel, Jamie E. DeNizio, Xiaohui Yan, Edward Chuang, Amber Tariq, Ryan R. Cupo, Laura M. Castellano, Kim A. Caldwell, Guy A. Caldwell, James Shorter

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Hsp104 is an AAA+ protein disaggregase that solubilizes and reactivates proteins trapped in aggregated states. We have engineered potentiated Hsp104 variants to mitigate toxic misfolding of α-synuclein, TDP-43, and FUS implicated in fatal neurodegenerative disorders. Though potent disaggregases, these enhanced Hsp104 variants lack substrate specificity and can have unfavorable off-target effects. Here, to lessen off-target effects, we engineer substrate-specific Hsp104 variants. By altering Hsp104 pore loops that engage substrate, we disambiguate Hsp104 variants that selectively suppress α-synuclein toxicity but not TDP-43 or FUS toxicity. Remarkably, α-synuclein-specific Hsp104 variants emerge that mitigate α-synuclein toxicity via distinct ATPase-dependent mechanisms involving α-synuclein disaggregation or detoxification of soluble α-synuclein conformers. Importantly, both types of α-synuclein-specific Hsp104 variant reduce dopaminergic neurodegeneration in a C. elegans model of Parkinson's disease more effectively than non-specific variants. We suggest that increasing the substrate specificity of enhanced disaggregases could be applied broadly to tailor therapeutics for neurodegenerative disease.

Original languageEnglish (US)
Pages (from-to)3314-3332.e9
JournalMolecular cell
Volume83
Issue number18
DOIs
StatePublished - Sep 21 2023

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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