Malate dehydrogenase from the extreme halophilic archaebacterium Halobacterium marismortui. Reconstitution of the enzyme after denaturation and dissociation in various denaturants

Malate dehydrogenase from the extreme halophilic archaebacterium Halobacterium marismortui is a homodimer of 84,000 molecular weight. As taken from ultracentrifugal analysis, it does not show concentration-dependent dissociation down to enzyme concentrations as low as 10 ng/mL. There is no change in...

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Bibliographic Details
Published in:Biochemistry (Easton) 1989-06, Vol.28 (12), p.4979-4985
Main Authors: Hecht, Katrin, Jaenicke, Rainer
Format: Article
Language:English
Subjects:
Bacteriology
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
malate dehydrogenase
Metabolism. Enzymes
Microbiology
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Summary:Malate dehydrogenase from the extreme halophilic archaebacterium Halobacterium marismortui is a homodimer of 84,000 molecular weight. As taken from ultracentrifugal analysis, it does not show concentration-dependent dissociation down to enzyme concentrations as low as 10 ng/mL. There is no change in specific activity at concentrations between 1 and 500 ng/mL. Deactivation is determined by the total ionic strength: keeping the salt concentration over the whole transition range beyond 4 M, the equilibrium transition occurs at 2.3 M Gdn multiplied by HCl (2.4 M NaCl). Reactivation after guanidine denaturation follows the same mechanism as described for the reconstitution after denaturation at low salt concentration. Low urea concentrations ( less than or equal to 2 M), as well as short incubation at low salt, lead to an increase in the yield of reconstitution due to incomplete dissociation under unfolding conditions.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi00438a012