Related Experiment Video
Updated: May 28, 2025
07:17
Purification of Hsp104, a Protein Disaggregase
Published on: September 30, 2011
17.3K
De novo designed Hsp70 activator dissolves intracellular condensates
Jason Z Zhang1, Nathan Greenwood2, Jason Hernandez3
1Department of Biochemistry, University of Washington, Seattle, WA 98195, USA; Institute for Protein Design, University of Washington, Seattle, WA 98195, USA; Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA.
Cell Chemical Biology
|February 8, 2025
Summary
Researchers designed novel Hsp70-binding proteins to control protein quality. These engineered proteins modulate Hsp70 activity, aiding in protein refolding and regulating cellular condensates, offering new tools for PQC research.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Protein quality control (PQC) relies on chaperones like Hsp70 and J-domain proteins (JDPs/Hsp40s).
- JDPs are believed to recruit Hsp70 to specific client proteins, but the underlying molecular mechanisms remain unclear.
Purpose of the Study:
- To design novel Hsp70-binding proteins that modulate Hsp70's ATPase activity.
- To investigate the functional consequences of engineered Hsp70 modulators in vitro and in vivo.
Main Methods:
- De novo design of Hsp70-binding proteins with inhibitory or stimulatory effects on ATPase activity.
- In vitro refolding assays using denatured luciferase.
- Targeting designed proteins to intracellular condensates in living cells.
Main Results:
- An ATPase-stimulating design successfully promoted in vitro refolding of denatured luciferase, mimicking native JDPs.
- Targeting this design to intracellular condensates led to their significant dissolution.
- The study revealed roles for these condensates as hubs promoting cell growth and signaling.
Conclusions:
- Engineered Hsp70-binding proteins provide insights into chaperone structure-function relationships.
- These designs offer a modular strategy to target PQC systems and regulate cellular condensates.
- The findings present new tools for manipulating protein quality control and cellular organization.
