The denaturation-renaturation of chicken-muscle triosephosphate isomerase in guanidinium chloride
| Autor: | Arthur Peacocke, John D. McVITTIE, M. Peter Esnouf |
|---|---|
| Rok vydání: | 1977 |
| Předmět: |
Guanidinium chloride
chemistry.chemical_classification Protein Denaturation Chromatography Protein Conformation Intrinsic viscosity Muscles Biochemistry Guanidines Triosephosphate isomerase Sedimentation coefficient Molecular Weight chemistry.chemical_compound Kinetics Monomer Enzyme chemistry Sedimentation equilibrium Animals Denaturation (biochemistry) Carbohydrate Epimerases Chickens Mathematics Triose-Phosphate Isomerase |
| Zdroj: | European journal of biochemistry. 81(2) |
| ISSN: | 0014-2956 |
| Popis: | 1. The process of denaturation of the chicken muscle dimeric enzyme triosephosphate isomerase on addition of guanidinium chloride has been studied at pH 7.6, the pH at which the recovery of activity is optimal (100%) on removal of denaturant. Determinations of the sedimentation coefficient, intrinsic viscosity, molecular weight (by sedimentation equilibrium studies) and the absorption coefficient at 280 nm in various concentrations of guanidinium chloride concurred in showing a single, sharp transition at about 0.7 M guanidinium chloride at a protein concentration 1-5 mg/ml from the native enzyme to the dissociated, unfolded chains of the monomer. Relative fluorescent intensity measurements revealed a single transition at about 0.4 M guanidinium chloride at enzyme concentrations of about 0.05 mg/ml. 2. The process of denaturation in different guanidinium chloride concentrations was first order with respect to enzyme and about sixth order with respect to denaturant. 3. The rate of attainment of equilibrium during the renaturation obeyed second-order/first-order reversible kinetics. It was concluded that the rate-determining step in renaturation at pH 7.6 must be the association of two subunits. |
| Databáze: | OpenAIRE |
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