Effect of temperature upon catalytic properties of lactate dehydrogenase isoenzymes from a poikilotherm

doi:10.1016/0005-2744(73)90254-4

Biochimica et Biophysica Acta (BBA) - Enzymology

Volume 315, Issue 2, 16 August 1973, Pages 250-258

https://doi.org/10.1016/0005-2744(73)90254-4 Get rights and content

Abstract

Lactate dehydrogenase isoenzymes A₄ and B₄ (l-lactate:NAD⁺ oxidoreductase, EC 1.1.1.27) have been purified from muscle and heartof the snake Bothrops neuwiedii. Catalytic properties at different temperatures and pH values have been studied compartively with those of the same isoenzymes from beef.

Values of $K_{m}$ for ophidian isoenzymes were markedly reduced when the temperature decreased. At low substrate concentrations, the increased enzyme-pyruvate affinity compensates for the lower thermal energy and the reaction rate appears to be independent of temperature. Under the same conditions, the activity of beef isoenzymes was closely related to thermal energy.

These findings suggest that lactate dehydrogenases from the poikilotherm are able to function with the same efficiency at all temperatures within the range of habitat and to adjust immediately to thermal changes.

References (17)

P.W. Hochachka et al.
Comp. Biochem. Physiol.
(1968)
P.W. Hochachka et al.
J. Biol. Chem.
(1970)
A.D. Winer et al.
J. Biol. Chem.
(1958)
J.A. Mayerle et al.
Comp. Biochem. Physiol.
(1971)
T. Moon et al.
Biochem. J.
(1971)
J. Baldwin et al.
Biochem. J.
(1970)
G.N. Somero et al.
Nature
(1969)
P.W. Hochchka et al.
Comp. Biochem. Physiol.
(1971)

There are more references available in the full text version of this article.

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In the present study, we demonstrated that the attachment of the nonpolar isopropylic carbon chain in the nitrogen of oxamate, converted this competitive inhibitor of LDH isozymes into a powerful selective inhibitor of mouse LDH-C4. The comparative study of the inhibitory effect of oxamate and N-isopropyl oxamate on mouse LDH isozymes pointed out that the isopropylic carbon chain conferred upon N-isopropyl oxamate a high affinity for LDH-C4 and a marked decrease in the affinity for the other isozymes since oxamate showed more inhibitory effect on LDH-1 (K_i=0.06 mM) and LDH-5 (K_i=0.08 mM), and less inhibitory effect on LDH-C4 (K_i=0.25 mM). On the other hand, N-isopropyl oxamate showed the highest inhibitory effect on LDH-C4 (K_i=0.014 mM) and poor inhibitory effect on LDH-1 (K_i=0.4 mM) and LDH-5 (K_i=0.8 mM). Apparently, the enzymatic inactivation proceeded through a reversible binding of N-isopropyl oxamate, facilitated by nonpolar interactions with a hydrophobic region present only in the active site of mouse LDH-C4, resulting in a selective inhibition of this isozyme in comparison with the other LDH isozymes. N-isopropyl oxamate was also a powerful competitive inhibitor of LDH-C4 (K_i=0.015 mM) compared with oxamate (K_i=0.35 mM), using α-ketoisocaproate as a substrate.
The effects of temperature and pH on the kinetic properties of heart muscle lactate dehydrogenase from anuran amphibians
1991, Comparative Biochemistry and Physiology -- Part B: Biochemistry and
- 1.
  1. The response to variation in pH and temperature of heart LDH enzyme from two anuran amphibians, Rana perezi and Bufo calamita, of different behaviour and habitat is studied.
- 2.
  2. Substrate (pyruvate and lactate) concentrations are measured in hearts from animals at rest and after exercise. In both species, substrate concentrations are higher than apparent K_m values of heart enzyme.
- 3.
  3. At physiological concentrations of substrate, pyruvate reductase activity is more affective by temperature increase when pH decreases simultaneously, whilst lactate oxidase reaction behaves conversely.
- 4.
  4. The contribution of cardiac metabolism to supporting the exercise effort in both species of anuran amphibians is discussed.
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1986, International Journal of Biochemistry
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  1. The temperature dependence of the kinetics of the yeast AM P deaminase was examined using the purified enzyme and the permeabilized yeast cells.
- 2.
  2. The increase in the enzyme affinity for the substrate AMP was accompanied by the decrease in the maximal velocity with the decreasing temperature in the absence and presence of ATP.
- 3.
  3. The apparent Km for AMP was lowest at 15–20°C, and the affinity was decreased below and above this temperature.
- 4.
  4. The rate of the AMP deaminase reaction remained constant over a wide range of temperature in the presence of physiological AMP concentrations.
- 5.
  5. The temperature dependent change in kinetic properties of AMP deaminase may contribute to the control of the yeast glycolytic flux under the condition of lower temperature environments.
Temperature and pH effects on the total white muscle LDH of Oreochromis niloticus (Pisces: Cichlidae)
1986, Comparative Biochemistry and Physiology -- Part B: Biochemistry and
- 1.
  1. The total lactate dehydrogenase enzyme activity of white muscle from O. niloticus shows positive thermal modulation.
- 2.
  2. There is a tendency towards conservation of binding capacity for substrate at physiologically changing pH conditions.
- 3.
  3. Inhibition by excess pyruvate is influenced by temperature and pH, but this phenomenon is not considered to be important in anaerobic glycolysis.
Effect of temperature upon catalytic properties of α-hydroxyacid dehydrogenase from Trypanosoma cruzi
1984, Molecular and Biochemical Parasitology
A comparative study of the effect of temperature (10, 20, 30 and 37°C) upon K_m and V of α-hydroxyacid dehydrogenase (HADH), isozyme I and II, from Trypanosoma cruzi, a parasite whose life cycle comprises stages in an insect vector, and of another enzyme with analogous substrate specificity, the lactate dehydrogenase, isozyme X (LDH X) from mouse, a homeotherm, is presented. The K_m for α-ketoisocaproate of HADH is markedly reduced as temperature decreases. This effect can compensate the reduction in thermal energy and produce stabilization of the reaction rate. This compensation does not occur with mouse LDH X. The activation energy for both HADH isozymes is about half the value determined for mouse LDH X. Results indicate that HADH from T. cruzi is able to adjust instantaneously to thermal changes of the environment, behaving as other enzymes of terrestrial poikilothermic animals.
H4-isozyme of lactate dehydrogenase in the solution of sodium chloride-3. The enzymatic activity and the pyruvate inhibition
1983, International Journal of Biochemistry
- 1.
  1. Low enzymatic activities in low pyruvate concentrations and high K_m were observed in sodium chloride solutions.
- 2.
  2. The pyruvate inhibition shown by the % activity at 1 mM pyruvate was lower in 0.5 M sodium chloride than in 0.1 M sodium phosphate.
- 3.
  3. At 40 C. as compared with results at 20 C, less pyruvate inhibition was observed in phosphate buffer and in sodium chloride solutions.
- 4.
  4. By using the equilibrium constants between dimer and tetramer, a theoretical explanation is proposed for the pyruvate inhibition. In this explanation, it is suggested that the quaternary complex which is composed of tetrameric enzyme, coenzyme and two kinds of pyruvates was the main cause of the pyruvate inhibition.

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Effect of temperature upon catalytic properties of lactate dehydrogenase isoenzymes from a poikilotherm

Abstract

Comp. Biochem. Physiol.

J. Biol. Chem.

J. Biol. Chem.

Comp. Biochem. Physiol.

Biochem. J.

Biochem. J.

Nature

Comp. Biochem. Physiol.