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Purification and partial characterization of malate dehydrogenase (decarboxylating) from Tritrichomonas foetus hydrogenosomes

Published online by Cambridge University Press:  06 April 2009

I. hrdý  and
E. Mertens
I. hrdý
Affiliation:
Department of Parasitology, Charles University, Prague, Czech Republic
E. Mertens
Affiliation:
Laboratoire de Chimie Physiologique, Universite Catholique de Louvain and International Institute of Cellular and Molecular Pathology, Brussels, Belgium
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Summary

Malate dehydrogenase (decarboxylating) from Tritrichomonas foetus hydrogenosomes was purified close to homogeneity by a combination of differential centrifugation, zwitterionic detergent solubilization, Red-Sepharose chromatography and anion-exchange chromatography. The enzyme with apparent subunit size of 59 kDa and native molecular mass of 308 kDa utilized NAD+ preferentially to NADP+ as a cofactor and required Mn2+ or Mg2+ for its activity. Affinity curves for malate and coenzymes were hyperbolic. Km for malate was 100 μM and 458 μM in the presence of NAD+ and NADP+, respectively. Km for NAD+ and for NADP+ in the presence of malate was 18 μM and 207 μM, respectively. The enzyme is proposed to be a tetramer with a possible physiological role in the maintenance of an appropriate NAD+/NADH ratio in hydrogenosomes.

Keywords

Tritrichomonas foetustrichomonadshydrogenosomesmalic enzyme
Type
Research Article
Information
Parasitology , Volume 107 , Issue 4 , November 1993 , pp. 379 - 385
Copyright
Copyright © Cambridge University Press 1993

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