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Updated: Feb 28, 2026

Isolating Potentiated Hsp104 Variants Using Yeast Proteinopathy Models

Isolating Potentiated Hsp104 Variants Using Yeast Proteinopathy Models

Published on: November 11, 2014

Substrate Discrimination by ClpB and Hsp104.

Danielle M Johnston¹, Marika Miot¹, Joel R Hoskins¹

¹Laboratory of Molecular Biology, National Cancer Institute, National Institutes of HealthBethesda, MD, United States.

Frontiers in Molecular Biosciences

|June 15, 2017

Summary

ClpB and Hsp104 molecular chaperones disaggregate proteins. Their substrate specificity is determined by nucleotide binding domain-1, even without the Hsp70 system.

Keywords:

ClpB DnaK Hsp104 Hsp70 aggregate amyloid disaggregase molecular chaperone

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Isolating Potentiated Hsp104 Variants Using Yeast Proteinopathy Models

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Purification of Hsp104, a Protein Disaggregase

Purification of Hsp104, a Protein Disaggregase

Published on: September 30, 2011

Area of Science:

Molecular biology
Biochemistry
Cell biology

Background:

ClpB (E. coli) and Hsp104 (yeast) are homologous AAA+ chaperones essential for thermotolerance.
They disaggregate and reactivate proteins aggregated during cellular stress, collaborating with Hsp70 systems.
Hsp104 is crucial for yeast prion propagation; protein aggregation links to diseases like neurodegeneration and cancer.

Purpose of the Study:

To investigate the intrinsic substrate preferences of ClpB and Hsp104.
To determine the role of the Hsp70 chaperone system in ClpB/Hsp104 substrate specificity.
To elucidate the molecular determinants of ClpB and Hsp104 substrate recognition.

Main Methods:

In vitro assays measuring protein disaggregation activity.
Comparative analysis of ClpB and Hsp104 activity in the presence and absence of Hsp70.
Biochemical characterization of ClpB and Hsp104 domains.

Main Results:

ClpB and Hsp104 exhibit distinct substrate preferences even without Hsp70 assistance.
Nucleotide binding domain-1 (NBD1) plays a critical role in determining substrate specificity for both chaperones.
The absence of Hsp70 does not abolish the innate substrate recognition capabilities of ClpB and Hsp104.

Conclusions:

Substrate specificity of ClpB and Hsp104 is primarily dictated by NBD1.
These chaperones possess intrinsic recognition mechanisms independent of Hsp70.
Understanding these mechanisms advances knowledge of protein quality control and disease-related protein aggregation.