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Purification and properties of phosphoenolpyruvate carboxylase from Molinema dessetae (Nematoda: Filarioidea)

Published online by Cambridge University Press:  06 April 2009

P. M. Loiseau ,
P. Gayral  and
F. Petek
P. M. Loiseau*
Affiliation:
Biologie et Contrôle des Organismes Parasites, Université de Paris-Sud, 92296- Châtenay-Malabry Cédex
P. Gayral
Affiliation:
Biologie et Contrôle des Organismes Parasites, Université de Paris-Sud, 92296- Châtenay-Malabry Cédex
F. Petek
Affiliation:
Régulation de l’Expression des Gènes, U.P.R. 37 C.N.R.S., Institut de Recherches Scientifiques sur le Cancer, 94802-Veillejuif Cédex
*
*Reprint requests to Dr P. M. Loiseau
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Summary

The presence of phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31), an enzyme at the branchpoint of glycolysis and the Krebs cycle was detected in the Filaria Molinema dessetae. This enzyme has not previously been identified in Helminths, which have so far been found to only possess a phosphoenolpyruvate carboxykinase (EC 4.1.1.32). This enzyme had a level of activity comparable to that of pyruvate kinase, and was relatively less active than enzymes such as malate dehydrogenase or lactate dehydrogenase. We propose here a method of purification of M. dessetae PEP-carboxylase. When purified to electrophoretic homogeneity, the enzyme had a molecular weight of 64 kDa. Kinetic studies indicated that the carboxylation reaction had an optimal pH of 5·8. The enzyme was inhibited by cations such as Fe2+, Zn2+, Cd2+, Cu2+ but required the presence of Mg2+ or Mn2+. The enzyme was thermostable. The apparent Km value of 2·38 mmol for phosphoenolpyruvate for the carboxylation reaction was higher than previously reported values. The Km value for KHCO3 was found to be 1·6 mmol. PEP-carboxylase did not catalyse the reverse reaction.

Keywords

phosphoenolpyruvate carboxylaseMolinema dessetaeFilarioideaFilaria.
Type
Research Article
Information
Parasitology , Volume 106 , Issue 1 , January 1993 , pp. 55 - 61
Copyright
Copyright © Cambridge University Press 1993

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