Hsp104 Disaggregase
Hsp104 Disaggregase - Related Videos
07:17
Purification of Hsp104, a Protein Disaggregase
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Cited by 24 •
2011
Here, we describe a protocol for the purification of highly active Hsp104, a hexameric AAA+ protein from yeast, which couples ATP hydrolysis to protein disaggregation. This scheme exploits a His6-tagged construct for affinity purification from E. coli followed by anion-exchange chromatography, His6-tag removal with TEV protease, and size-exclusion chromatography.
08:44
Isolating Potentiated Hsp104 Variants Using Yeast Proteinopathy Models
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Cited by 16 •
2014
Yeast proteinopathy models are valuable tools to assess the toxicity and aggregation of proteins implicated in disease. Here, we present methods for screening Hsp104 variant libraries for toxicity suppressors. This protocol could be adapted to screen any protein library for toxicity suppressors of any protein that is toxic in yeast.
08:12
High-throughput Screening for Protein-based Inheritance in S. cerevisiae
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Cited by 2 •
2017
This protocol describes a high-throughput methodology to functionally screen for protein-based inheritance in S. cerevisiae.
FAQs
The six-subunit ring creates a central pore through which an aggregated polypeptide can be drawn. This arrangement couples substrate engagement at the pore with ATP-dependent mechanical activity, allowing Hsp104 to act on proteins trapped in disordered or amyloid-like assemblies. The ring architecture is therefore essential for converting molecular energy into movement that helps separate aggregated protein material.
ATP hydrolysis supplies the energy required to pull polypeptide segments through Hsp104’s central pore. Rather than merely binding an aggregate, the AAA+ ATPase uses repeated nucleotide-driven activity to remodel its protein substrate. This energy dependence explains how the complex can help restore proteins from physically stable assemblies and support recovery from proteotoxic stress.
Hsp70 and Hsp40 can work with Hsp104 to remodel aggregated proteins. Their cooperation adds chaperone support to the disaggregation process, helping Hsp104 act on substrates that may be disordered or amyloid-like. This partnership connects aggregate dismantling with broader cellular proteostasis, the system that maintains proteins in suitable functional states.
Hsp104 can remodel both disordered aggregates and amyloid-like assemblies, which differ in their organization and stability. Its pore-based, ATP-dependent mechanism provides a way to engage and pull polypeptide material from these structures rather than treating every aggregate as identical. This breadth makes the disaggregase relevant to diverse forms of protein-misfolding research.
During heat shock or other damaging conditions, proteins can accumulate in toxic aggregates that disrupt proteostasis. Hsp104 contributes to recovery by dismantling those assemblies and helping restore soluble, functional protein. Studying this response shows how cells use molecular chaperone systems not only to withstand proteotoxic stress, but also to regain protein function afterward.
Hsp104 provides a model for examining how protein aggregates form, persist, and can be remodeled. Its ability to act on amyloid-like assemblies connects the system to prion biology, while aggregate toxicity and protein misfolding are central concerns in neurodegenerative disease research. Experiments involving Hsp104 can therefore link basic proteostasis mechanisms with disease-related aggregation.
Engineered Hsp104 variants are being investigated because modified disaggregase activity could offer new ways to address harmful protein assemblies. These studies have therapeutic relevance for conditions involving protein misfolding and may also support biotechnology applications that require proteins to remain soluble or recover function. The work extends Hsp104 research beyond natural cellular stress responses.