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Macromolecular crowding perturbs protein refolding kinetics: implications for folding inside the cell
B van den Berg1, R Wain, C M Dobson
1Oxford Centre for Molecular Sciences, New Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QT, UK.
The EMBO Journal
|August 2, 2000
Summary
Macromolecular crowding affects protein folding kinetics. While folding energetics remain similar, crowding accelerates fast folding tracks and retards slow tracks, mimicking cellular environments.
Area of Science:
- Biochemistry
- Physical Chemistry
- Molecular Biology
Background:
- Macromolecular crowding is a ubiquitous feature of the cellular environment.
- Understanding its impact on protein folding is crucial for cell biology and drug design.
- Hen lysozyme is a model protein for studying folding pathways.
Purpose of the Study:
- To investigate the influence of macromolecular crowding on the kinetics of hen lysozyme refolding.
- To determine if crowding alters the fundamental energetics of protein folding.
- To elucidate the specific effects of crowding agents like bovine serum albumin and Ficoll 70 on folding pathways.
Main Methods:
- Oxidative refolding of hen lysozyme.
- Comparison of refolding kinetics in dilute solution versus high concentrations of bovine serum albumin and Ficoll 70.
- Analysis of refolding intermediates and their accumulation levels.
Main Results:
- The heterogeneity of lysozyme refolding is maintained under crowded conditions.
- Refolding intermediates show similar accumulation in the presence and absence of crowding agents.
- Crowding accelerates the fast folding pathway but significantly retards the slow folding pathway.
- Results suggest preferential stabilization of compact states due to excluded volume effects.
Conclusions:
- Macromolecular crowding does not substantially alter the energetics of hen lysozyme folding.
- Crowding significantly modifies the kinetics of different folding pathways.
- Protein folding rates in vivo may differ substantially from those observed in dilute solutions due to crowding effects.