Specific volume and adiabatic compressibility measurements of native and aggregated recombinant human interleukin-1 receptor antagonist: Density differences enable pressure-modulated refolding
| Autor: | Brent S. Kendrick, Matthew B. Seefeldt, Chris Crouch, Theodore W. Randolph |
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| Rok vydání: | 2007 |
| Předmět: |
Protein Folding
Chemistry Molecular Conformation Bioengineering Protein aggregation Applied Microbiology and Biotechnology Recombinant Proteins law.invention Interleukin 1 Receptor Antagonist Protein Structure-Activity Relationship Crystallography Volume (thermodynamics) law Partial specific volume Yield (chemistry) Pressure Native state Recombinant DNA Biophysics Humans Thermodynamics Protein folding Biotechnology Bar (unit) |
| Zdroj: | Biotechnology and Bioengineering. 98:476-485 |
| ISSN: | 1097-0290 0006-3592 |
| DOI: | 10.1002/bit.21398 |
| Popis: | High hydrostatic pressures have been used to dissociate non-native protein aggregates and foster refolding to the native conformation. In this study, partial specific volume and adiabatic compressibility measurements were used to examine the volumetric contributions to pressure-modulated refolding. The thermodynamics of pressure-modulated refolding from non-native aggregates of recombinant human interleukin-1 receptor antagonist (IL-1ra) were determined by partial specific volume and adiabatic compressibility measurements. Aggregates of IL-1ra formed at elevated temperatures (55 degrees C) were found to be less dense than native IL-1ra and refolded at 31 degrees C under 1,500 bar pressure with a yield of 57%. Partial specific adiabatic compressibility measurements suggest that the formation of solvent-free cavities within the interior of IL-1ra aggregates cause the apparent increase in specific volume. Dense, pressure-stable aggregates could be formed at 2,000 bar which could not be refolded with additional high pressure treatment, demonstrating that aggregate formation conditions and structure dictate pressure-modulated refolding yields. |
| Databáze: | OpenAIRE |
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