High Pressure Fosters Protein Refolding from Aggregates at High Concentrations

High hydrostatic pressures (1-2 kbar), combined with low, nondenaturing concentrations of guanidine hydrochloride (GdmHCl) foster disaggregation and refolding of denatured and aggregated human growth hormone and lysozyme, and β -lactamase inclusion bodies. One hundred percent recovery of properly fo...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1999-11, Vol.96 (23), p.13029-13033
Main Authors: St. John, Richard J., Carpenter, John F., Randolph, Theodore W.
Format: Article
Language:English
Subjects:
Aggregates
Aggregation
Atmospheric pressure
beta-Lactamases - chemistry
Biochemistry
Biological Sciences
Cell aggregates
Disulfides
Growth Hormone - chemistry
Guanidine - chemistry
High pressure
Hormones
Humans
Inclusion bodies
Molecular biology
Molecules
Muramidase - chemistry
Pressure
Protein Denaturation
Protein Folding
Protein refolding
Proteins
Recombinant Proteins - chemistry
Water pressure
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Summary:High hydrostatic pressures (1-2 kbar), combined with low, nondenaturing concentrations of guanidine hydrochloride (GdmHCl) foster disaggregation and refolding of denatured and aggregated human growth hormone and lysozyme, and β -lactamase inclusion bodies. One hundred percent recovery of properly folded protein can be obtained by applying pressures of 2 kbar to suspensions containing aggregates of recombinant human growth hormone (up to 8.7 mg/ml) and 0.75 M GdmHCl. Covalently crosslinked, insoluble aggregates of lysozyme could be refolded to native, functional protein at a 70% yield, independent of protein concentration up to 2 mg/ml. Inclusion bodies containing β -lactamase could be refolded at high yields of active protein, even without added GdmHCl.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.96.23.13029