Reduction of Cys36-Cys42 and Cys64-Cys74 disulfide bonds in recombinant human granulocyte colony stimulating factor
| Autor: | Auke Bult, J. Leon, E. Reubsaet, Willy J. M. Underberg, Arjan J. Hoekstra, Jan Teeuwsen, Jos H. Beijnen, Peter J.H.J. van Os, Wout P. van Bennekom, Ed Hop |
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| Rok vydání: | 2000 |
| Předmět: |
Stereochemistry
Clinical Biochemistry Kinetics Molecular Sequence Data Pharmaceutical Science Granulocyte Redox Medicinal chemistry Dithiothreitol Mass Spectrometry Analytical Chemistry law.invention Catalysis chemistry.chemical_compound symbols.namesake law Drug Discovery Granulocyte Colony-Stimulating Factor medicine Humans Nernst equation Amino Acid Sequence Cysteine Disulfides Spectroscopy Chromatography High Pressure Liquid Chemistry Hydrogen-Ion Concentration Recombinant Proteins Granulocyte colony-stimulating factor medicine.anatomical_structure Recombinant DNA symbols Oxidation-Reduction Chromatography Liquid |
| Zdroj: | Journal of pharmaceutical and biomedical analysis. 19(6) |
| ISSN: | 0731-7085 |
| Popis: | The Cys36-Cys42 and Cys64-Cys74 disulfide bonds in recombinant methionyl human granulocyte colony-stimulating factor were reduced to sulfhydryls with dithiothreitol or mercury. Both reduction reactions are dependent on the pH. The reduction reaction with dithiothreitol increased in rate with increasing pH; between pH 7-9 and above pH 10.5 this increase was less than in other regions. These observations are explained by repulsive forces between dithiothreitol and regions in granulocyte colony-stimulating factor which intensify in these pH-regions. The hydroxyl catalysis causes the overall increase in k(obs) in the pH-region studied. The reduction of the disulfides with mercury is, as could be expected from the Nernst equation for disulfide reduction, also pH dependent: the half-wave potential decreases with increasing pH as predicted by theory. |
| Databáze: | OpenAIRE |
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