Counteraction of trehalose on urea-induced protein unfolding: Thermodynamic and kinetic studies |
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| Affiliation: | 1. Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary;2. Department of Biophysics and Radiation Biology, Semmelweis University, MTA-SE Molecular Biophysics Research Group, Budapest, Hungary;1. Department of Cell Signaling, Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119992 Russia;2. Protein Research Group, Institute for Biological Instrumentation of the Russian Academy of Sciences, Pushchino, Moscow region, 142290 Russia;3. Department of Neurosciences, Biochemistry Group, University of Oldenburg, Oldenburg, 26111 Germany;4. Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Pushchino, Moscow region, 142290 Russia;5. Department of Life Sciences and Reproduction, Section of Biological Chemistry and Center for BioMedical Computing, University of Verona, Verona, 37134 Italy;1. Department of Chemistry, National Institute of Technology, Meghalaya, Shillong 793003, India;2. Centre for Advanced Studies, Department of Chemistry, North-Eastern Hill University, Shillong 793022, India |
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| Abstract: | Urea-induced protein denaturation can be effectively inhibited by trehalose, but the thermodynamic and kinetic behaviors are still unclear. Herein, the counteraction of trehalose on urea-induced unfolding of ferricytochrome c was studied. Thermodynamic parameters for the counteraction of trehalose were derived based on fluorescence spectroscopic data. Then the kinetics was emphatically investigated by stopped-flow fluorescence spectroscopy. Urea-induced unfolding of ferricytochrome c in 8.00 mol/L urea solution reveals two observable phases, including fast and slow phases following a burst phase. Trehalose has little influence on the burst phase amplitude. Nevertheless, the observable unfolding pathway is significantly affected by trehalose. At lower trehalose concentrations (<0.20 mol/L) in 8.00 mol/L urea, the unfolding pathways still keep to show two phases. However, the rate constant and amplitude for the fast phase diminish with increasing trehalose concentration. In contrast, the rate constant for the slow phase shows only a slight change with a significant increase of the amplitude. At higher trehalose concentrations (>0.30 mol/L), the unfolding pathway is transformed into a single slow phase. The rate constant and amplitude for the single phase also decrease with increasing trehalose concentration. The studies are expected to help our understanding of trehalose effects on protein stability. |
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| Keywords: | Protein Biophysical chemistry Protein denaturation Kinetic parameters Trehalose Urea |
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