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Coupled Assays for Monitoring Protein Refolding in Saccharomyces cerevisiae
Published on: July 9, 2013
Hyperthermophilic archaeal prefoldin shows refolding activity at low temperature
Tamotsu Zako1, Shinya Banba, Muhamad Sahlan
1Bioengineering Laboratory, RIKEN Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. zako@riken.jp
Biochemical and Biophysical Research Communications
|November 19, 2009
Summary
This study reveals that Pyrococcus horikoshii OT3 prefoldin (PhPFD) can refold denatured lysozyme. This molecular chaperone exhibits refolding activity at lower temperatures, challenging previous understanding of prefoldin function.
Area of Science:
- Molecular Biology
- Biochemistry
- Protein Folding
Background:
- Prefoldin acts as a molecular chaperone, capturing folding intermediates for transfer to chaperonins.
- Previously, archaeal prefoldins were understood to possess only holdase activity, lacking intrinsic refolding capabilities.
Purpose of the Study:
- To investigate the refolding activity of a prefoldin from the hyperthermophilic archaeon Pyrococcus horikoshii OT3 (PhPFD).
- To characterize the interaction between PhPFD and denatured lysozyme at varying temperatures.
Main Methods:
- Denatured lysozyme refolding assays.
- Surface Plasmon Resonance (SPR) sensor analysis to study protein-protein interactions at different temperatures.
Main Results:
- PhPFD demonstrated refolding activity for denatured lysozyme at temperatures lower than typical physiological conditions.
- SPR analysis showed strong PhPFD-lysozyme affinity at high temperatures but weak affinity at lower temperatures.
- Protein folding appears to be facilitated by the dynamic binding and release of substrate mediated by this temperature-dependent affinity.
Conclusions:
- The prefoldin PhPFD exhibits intrinsic protein refolding capabilities, expanding the known functions of prefoldins.
- The temperature-dependent binding affinity of PhPFD is crucial for its refolding mechanism.
- Prefoldins may contribute to the folding of cellular proteins that exhibit weak interactions with the chaperone.
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