Effects of Low Concentrations of Guanidine . HCl on the Reconstitution of Lactic Dehydrogenase from Pig Muscle in vitro. Evidence for Guanidine Binding to the Native Enzyme
- 1 October 1979
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 100 (2), 593-598
- https://doi.org/10.1111/j.1432-1033.1979.tb04206.x
Abstract
The presence of low concentrations of guanidine.cntdot.HCl has a pronounced effect on the overall rate of reactivation of lactic dehydrogenase from pig muscle after preceding dissociation and deactivation in various denaturants. The observed attenuation is a function of the amount of guanidine.cntdot.HCl present during reconstitution. At a given guanidine concentration in the reactivation buffer the yield, but not the rate of reactivation, is influenced by the extent of denaturation caused initially in the process of deactivation and dissociation. As a possible explanation for the influence of guanidine.cntdot.HCl on the kinetics of reconstitution, binding of the ligand to intermediates of folding and association is considered. This hypothesis is corroborated by the observation that guanidine.cntdot.HCl in the relevant concentration range does bind to native lactic dehydrogenase without inactivating the enzyme or disrupting its quaternary structure. A kinetic model comprising guanidine binding to both the native enzyme and structural intermediates is proposed to describe the observed effects of guanidine.cntdot.HCl on the rate of reactivation. In addition, the Kd for guanidine binding to intermediates of reconstitution and to native lactic dehydrogenase are estimated.This publication has 10 references indexed in Scilit:
- Quaternary structure, subunit activity, and in vitro association of porcine mitochondrial malic dehydrogenaseBiochemistry, 1979
- Kinetics of in vitro reconstitution of oligomeric enzymes by cross-linkingNature, 1979
- Effect of Zinc(II) on the Refolding and Reactivation of Liver Alcohol DehydrogenaseEuropean Journal of Biochemistry, 1978
- Equilibrium Studies and Kinetics of Reactivation, Refolding and Reassociation of Lactic Dehydrogenase and Glyceraldehyde-3- Phosphate DehydrogenasePublished by Walter de Gruyter GmbH ,1977
- Effect of coenzymes and temperature on the process of in vitro refolding and reassociation of lactic dehydrogenase isoenzymesBiochemistry, 1977
- Mechanism of refolding and reactivation of lactic dehydrogenase from pig heart after dissociation in various solvent mediaBiochemistry, 1977
- kinetics of reactivation of rabbit muscle aldolase after denaturation and dissociation in various solvent mediaEuropean Biophysics Journal, 1977
- Kinetics of Reassociation and Reactivation of Pig-Muscle Lactic Dehydrogenase after Acid DissociationEuropean Journal of Biochemistry, 1976
- Molecular Weight and Quaternary Structure of Lactic DehydrogenaseEuropean Journal of Biochemistry, 1968
- Some Factors in the Interpretation of Protein DenaturationAdvances in protein chemistry, 1959