Cosolvent Assisted Protein Refolding
| Autor: | Daniel I. C. Wang, Jeffrey L. Cleland |
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| Rok vydání: | 1990 |
| Předmět: |
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Protein Conformation Kinetics Biomedical Engineering Bioengineering Polyethylene glycol Applied Microbiology and Biotechnology Polyethylene Glycols Structure-Activity Relationship chemistry.chemical_compound Protein structure Carbonic anhydrase PEG ratio Animals Scattering Radiation Structure–activity relationship Carbonic Anhydrases Aqueous solution biology Chemistry Solutions Biochemistry Protein refolding biology.protein Biophysics Molecular Medicine Cattle Biotechnology |
| Zdroj: | Nature Biotechnology. 8:1274-1278 |
| ISSN: | 1546-1696 1087-0156 |
| Popis: | The use of cosolvents in aqueous systems has been shown to enhance protein refolding and decrease aggregation. In this study, we have used polyethylene glycol (PEG) in the molecular weight range of 1000 to 8000 Daltons to effectively increase the rate of refolding and prevent aggregation of the model protein, bovine carbonic anhydrase B (CAB). At concentrations of 3 and 30 g/l, PEG increased the rate of recovery of active protein in the absence of aggregation. Using 3 g/l PEG (3350 MW), the refolding rate was three fold greater than the observed normal refolding rate. The observed rate enhancement was caused by PEG acting on the first intermediate in the CAB refolding pathway to increase the rate of formation of the second intermediate. The interaction of PEG with the first intermediate also prevented its self-association during refolding and at equilibrium. The stabilization of this first intermediate resulted in complete recovery of active protein under normal aggregating conditions. |
| Databáze: | OpenAIRE |
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