| | 1 | Author
| G. Ünther Bernhardt, Rainer Rudolph, Rainer Jaenicke | Requires cookie* | | | Title
| Reassociation of Lactic Dehydrogenase from Pig Heart Studied by Cross-Linking with Glutaraldehyde  | | | | Abstract
| Cross-linking with glutaraldehyde has been successfully applied in order to analyze the kinetics of reassociation of oligomeric enzymes (R. Hermann, R. Rudolph, and R. Jaenicke Nature 277, 243-245 (1979)). In the present study the assembly of lactic dehydrogenase from pig heart is investigated using this approach. In order to eliminate perturbations caused by excessive folding reactions, acid dissociation was performed in the presence of 0.8 M Na2S 0 4 at 0°C . Under optimum conditions complete cross-linking of the tetrameric enzyme was achieved in less than 2 minutes. Cross-linking during reconstitution proves the dimer to be the only intermediate of reasso ciation. The dimer -*■ tetramer transition is found to be rate-limiting for both reassociation and reactivation, suggesting the tetramer to be the enzymatically active species. The presence of monomers during reconstitution indicates that tetramer formation is preceded by a fast monomer-dimer equilibrium. The kinetic model describing the experimental data 4M^2D^T is characterized by an equilibrium constant K = 3 ± 1 • 107 liter • m ol-1, and a second-order rate constant k = 1.4 ± 0.2 • 104 liter • mol-1 • s-1. | | | |
Reference
| Z. Naturforsch. 36c, 772—777 (1981); received May 15 1981 | | | |
Published
| 1981 | | | |
Keywords
| Cross-Linking, Glutaraldehyde, Lactic Dehydrogenase, Reassociation, Reconstitution | | | |
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| | | | | TEI-XML for
| default:Reihe_C/36/ZNC-1981-36c-0772.pdf | | | | Identifier
| ZNC-1981-36c-0772 | | | | Volume
| 36 | |
| 2 | Author
| Axel Ganzhom, Margrit Scharschm, G.Erhard Pfleiderer | Requires cookie* | | | Title
| Über die pH-induzierte Inaktivierung der Glycerindehydrogenase aus Bacillus megaterium | | | | Abstract
| -Induced In activ atio n o f G ly cero l D e h y d ro g e n a s e from Bacillus m egaterium The kinetic o f the reversible dissociation o f glycerol dehydrogenase from Bacillus m egaterium in slightly alkaline medium was measured by biochem ical, chem ical and physical m ethods. The dissociation is followed by changes in the secondary structure and can be prevented by addition o f N A D or increased potassium chloride concentration. Crosslinking by suberim idate, but not by monofunctional im ido esters, shows a high stabilization against alkali, urea or heat inactivation caused by hindrance o f dissociation. | | | |
Reference
| Z. Naturforsch. 39c, 575—583 (1984); received January 31 1984 | | | |
Published
| 1984 | | | |
Keywords
| Dissociation, Reassociation, Glycerol Dehydrogenase, Crosslinking, Denaturation | | | |
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| default:Reihe_C/39/ZNC-1984-39c-0575.pdf | | | | Identifier
| ZNC-1984-39c-0575 | | | | Volume
| 39 | |
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