Susceptibility towards intramolecular disulphide-bond formation affects conformational stability and folding of human basic fibroblast growth factor
| Autor: | Ursula Rinas, J van den Heuvel, David Estapé |
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| Rok vydání: | 1998 |
| Předmět: |
Models
Molecular Protein Denaturation Protein Folding Protein Conformation Reducing agent Globular protein Biochemistry Mice Protein structure Cadmium Chloride Animals Humans Urea Molecule Denaturation (biochemistry) Cysteine Disulfides Molecular Biology chemistry.chemical_classification Chemistry 3T3 Cells Cell Biology Crystallography Spectrometry Fluorescence Intramolecular force Fibroblast Growth Factor 2 Protein folding Oxidation-Reduction Research Article |
| Zdroj: | Biochemical Journal. 335:343-349 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj3350343 |
| Popis: | The conformational stability and the folding properties of the all-beta-type protein human basic fibroblast growth factor (hFGF-2) were studied by means of fluorescence spectroscopy. The results show that the instability of the biological activity of hFGF-2 is also reflected in a low conformational stability of the molecule. The reversibility of the unfolding and refolding process was established under reducing conditions. Determination of the free-energy of unfolding in the presence of reducing agents revealed that the conformational stability of hFGF-2 (DeltaGH2Oapp congruent with21 kJ. mol-1, 25 degreesC) is low compared with other globular proteins under physiological conditions (20-60 kJ.mol-1). However, the conformational stability of hFGF-2 is particularly low under non-reducing conditions. This instability is attributed to intramolecular disulphide-bond formation, rendering the molecule more susceptible to denaturant-induced unfolding. In addition, denaturant-induced unfolding of hFGF-2 renders the protein more susceptible to irreversible oxidative denaturation. Experimental evidence is provided that the irreversibility of the unfolding and refolding process in the absence of reducing agents is linked to the formation of an intramolecular disulphide bond involving cysteines 96 and 101. |
| Databáze: | OpenAIRE |
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