Kinetic studies on pig heart cytoplasmic malate dehydrogenase.

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Kinetic studies on pig heart cytoplasmic malate dehydrogenase.

Kinetic studies on the pig heart cytoplasmic malate dehydrogenase have been performed over a wide range of conditions using the full time course of the reaction and computer simulation to obtain the kinetic parameters. The maximum velocity and Michaelis constants for the oxidation of reduced coenzyme have been determined as a fundtion of pH in 0.05 M phosphate buffer at 15 degrees. At pH 7.5 an...

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Kinetic studies on the mechanism of the malate dehydrogenase reaction.

This work is a kinetic investigation of the reaction mechanism of malate dehydrogenase, prepared from washed mince of whole bovine heart by a variation on previous methods. The forward and reverse reactions catalyzed by this enzyme have been studied at pH 8.0 in the presence and in the absence of one product at a time, with the use of a recording fluorometer to measure changes in the concentrat...

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Lifetimes and NADH quenching of tryptophan fluorescence in pig heart cytoplasmic malate dehydrogenase.

The time-resolved and steady state fluorescence properties were measured for pig heart cytoplasmic malate dehydrogenase at pH 6.0 and 8.0. The fluorescence decay can be described by two rate processes, according to the functions: I(t) = 0.7e(-t/1.0) + 0.3e(-t/4.4) for the free enzyme and I(t) = 0.7e(-t/0.8) + 0.3e(-t/2.0 for the enzyme . NADH complex. Quenching by NADH of the tryptophan fluores...

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Studies on the presence and role of tryptophan in pig heart mitochondrial malate dehydrogenase.

Several different experimental approaches have shown the presence of tryptophan in malate dehydrogenase from pig heart mitochondria. The methods used were (a) fluorescence measurement of tryptophan content after acid hydrolysis and separation of amino acids by paper chromatography, (b) recovery of tryptophan labeled with tritium after incubation of the enzyme with tritiated substrates, (c) dete...

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The folding of dimeric cytoplasmic malate dehydrogenase. Equilibrium and kinetic studies.

Porcine heart cytoplasmic malate dehydrogenase (s-MDH) is a dimeric protein (2 x 35 kDa). We have studied equilibrium unfolding and refolding of s-MDH using activity assay, fluorescence, far-UV and near-UV circular dichroism (CD) spectroscopy, hydrophobic probe-1-anilino-8-napthalene sulfonic acid binding, dynamic light scattering, and chromatographic (HPLC) techniques. The unfolding and refold...

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ژورنال

عنوان ژورنال: Journal of Biological Chemistry

سال: 1975

ISSN: 0021-9258

DOI: 10.1016/s0021-9258(19)41689-x