Properties of the energy-allowance enzymes of the common frog in different physiological body states. I. Adenosine triphosphatase and succinate dehydrogenase activities

The activity of SDG, Na, K-ATPase and Mg-ATPase of the grass frog was determined in January, March and May, the number of animals examined being 30-40 in either series of experiments. In May (period of reproduction) the average activity of the above enzymes was higher than in January and March. This was observed both in males and females. A correlation between enzymatic activities of each single organism in March and May significantly increased. The role of these changes in the increase of viability of the organism is discussed.     

Activity and thermal stability of the enzymes of energy support in the common frog

The activity and heat resistance of succinate dehydrogenase (SDG), Na, K-ATPase and Mg-ATPase of the grass frog (39 specimens) have been determined. No correlation was found between individual levels of heat resistance both of either enzyme examined and of the same enzyme but taken from different tissues (SDG of liver and muscles). The average level of heat resistance of SDG in liver is significantly higher than that in m. gastrocnemius. A statistically significant correlation was observed between individual levels of enzyme activity in the internal organs (SDG of liver, Na, K-ATPase and Mg-ATPase of kidney). The activity of SDG of muscles does not correlate with that of either partner.     

Physiological polymorphism of a population of the common frog. II. The polymorphism of the type of muscle tissue heat resistance in stress]

A study was made of the dynamics in the heat resistance of m. interphalangealis of individual grass frogs under stress. The response of muscle tissue to injury can be differentiated into three types according to the initial heat resistance level: increase, decrease and phase change of the resistance. The pattern of response of muscle tissue of individual frogs to stress is one of manifestationss of physiological polymorphism of the population.     

Adenosine triphosphatases associated with bovine brain microtubules. II. Properties of ATPases I and II

The catalytic properties of two ATPases which had been purified from bovine brain microtubules (Tominaga, S. & Kaziro, Y. (1983) J. Biochem. 93, 1085-1092) were studied. ATPase I, which had a molecular weight of 33,000, required the presence of 1.0 microM tubulin, 0.2 mM Mg2+, and 10 mM Ca2+ for maximal activity. The activation of ATPase I by tubulin was specific to the native form of tubulin, which could not be replaced by F-actin or tubulin denatured either by heat or more mildly by dialysis in the absence of glycerol. ATPase I was not specific to ATP, and GTP, and to a lesser extent, UTP and CTP were also hydrolyzed. Km for ATP of ATPase I was about 0.04 mM. ATPase I was inhibited by 5 mM Mg2+, 0.04 M K+, 10(-3) M vanadate, 10 mM N-ethylmaleimide, or 20% (v/v) glycerol. ATPase II, which was associated with membrane vesicles, required the presence of 0.2-2.0 mM Mg2+ and 20 mM KCl for activity. Tubulin stimulated the reaction of ATPase II only partially, and the addition of Ca2+ was rather inhibitory. ATPase II was specific to ATP with a Km value of 0.14 mM. It was inhibited by 1.6 mM N-ethylmaleimide and 20% (v/v) glycerol, but was not very sensitive to vanadate. Instead, ATPase II was inhibited by trifluoperazine, chlorpromazine, and nicardipin at 10(-3) M.     

Physiological polymorphism in a population of the common frog. I. The polymorphism of a population of the common frog for the trait of muscle tissue heat resistance]

A study was made of heat resistance (36+/-0.2 degrees C) of m. interphalangealis of the third finger of a hind extremity in animals of one population of Rana temporaria L. living on the boundary between Leningrad region and Pskov region. Within this population, the existence of three groups of individuals differing in heat resistance levels of their m. interphalangealis is postulated. This conclusion is based on the distribution curve of heat resistance values and on the application of the probit-method. The distribution of the frequency of occurrence of individuals in these three groups follows Hardy-Weinberg's equation.     

Correlations of the individual levels of the activity and heat resistance of energy-support enzymes in the lake frog

A study was made of the correlation between the levels of the activity and heat resistance of Na,K-ATPase, Mg-ATPase and succinate dehydrogenase (SDG) in Rana ridibunda. A correlation was observed between the activities of two ATPases, and between either of them and the activity of liver SDG. The correlation coefficient between the heat resistance levels was statistically significant only in SDG of liver and muscles. The question whether the criterion of T50% inactivation may be good for a comparative appraisal of the heat resistance of enzymes is under discussion.     

Changes in polyamine-oxidizing capacity of peroxisomes under various physiological conditions in rats

Rat liver peroxisomal polyamine oxidase activity was determined under various physiological conditions by using the peroxidase method with phenol and 4-aminoantipyrine. N1-Acetylpolyamines such as N1-acetylspermine and N1-acetylspermidine were better substrates than the free polyamines. The polyamine oxidase activity in rat peroxisomes increased significantly when cell proliferation was high. The activity began to appear in fetal liver at the 16th approximately 18th day of pregnancy and peaked in neonatal liver on the first day (approx. 1.7-times higher than in adult liver). In regenerating rat liver, only polyamine oxidase activity among the peroxisomal enzymes tested was increased considerably 12 h after partial hepatectomy (approx. 2.8-fold over the control liver). Finally, the enzyme activity was significantly increased by administration of clofibrate, a peroxisome proliferator, which also causes hepatomegaly. In all cases, the increase in polyamine oxidase activity was not more than 3-fold. Since the level of polyamine oxidase activity in the normal liver is more than adequate in relation to the level of the substrates, the slight but significant increase under conditions of cell proliferation may have a role in modulating levels of polyamines in the proliferating liver tissue.     

The localization and properties of membrane adenosine triphosphatases in the guinea-pig placenta.

The distribution and properties of cytochemically demonstrable phosphatases in the near-term guinea-pig placenta were examined using a strontium capture technique for sodium- and potassium-dependent adenosine triphosphatase (Na+, K+-ATPase) and a lead capture technique for magnesium-dependent adenosine triphosphatase (Mg2+-ATPase). Localizations with the strontium technique in the presence of an alkaline phosphatase inhibitor were mainly on the syncytiotrophoblast plasma membranes; the reaction was potassium-dependent and ouabain-sensitive. Reaction product using the lead capture method was found on both trophoblast and endothelial cell plasma membranes and was independent of magnesium and insensitive to p-hydroxymercuribenzoate (POHMB), an inhibitor of membrane ATPases. However, a very large proportion of this reaction could be blocked by an alkaline phosphatase inhibitor. It is concluded that the strontium capture technique gave a reliable localization for Na+, K+-ATPase. However, the lead capture method mainly demonstrated alkaline phosphatase, and does not offer a useful approach to specific ATPase studies in this particular system.     

Variations of various enzymatic activities of the Krebs cycle (malate dehydrogenase, isocitrate dehydrogenase, succinate dehydrogenase) during experimental ischemic shock in the rat. Influence of adenosine 5' triphosphoric acid]

During the ischemic shock caused by the removal of tourniquets placed on the hind paws of the rat, a marked decrease in the enzyme activities of Krebs cycle yielding ATP (malate dehydrogenase, isocitrate dehydrogenase, succinate dehydrogenase) at the level of the gastrocnemius muscle and the liver, was observed together with a plasma increase of these enzymes. The intraperitoneal injection of ATP diminishes significantly the variations observed.     

Nature of the activation of succinate dehydrogenase by various effectors and in hypobaria and hypoxia

1. Hepatic mitochondrial succinate dehydrogenase (succinate:(acceptor)oxidoreductase, EC 1.3.99.1) was activated by preincubation of mitochondria with four diverse classes of compounds, the dicarboxylic acids, nitrophenols, quinols (and ubiquinols) and pyrophosphates. Of the various compounds tested malonate, oxaloacetate and pyrophosphate, well-known competitive inhibitors of the enzyme, and also hydroquinone and ubiquinols were effective even at low concentrations and showed maximal stimulation in 2 min.2. Activation of succinate dehydrogenase by ubiquinol-9 and ubiquinol-10 was comparable to succinate activation in fresh mitochondria, and was much higher in the aged samples.3. Preincubation of mitochondria with succinate, 2,4-dinitrophenol, pyrophosphate and ATP also stimulated the succinate-2,2′,5,5′-tetraphenyl-3,3′-(4,4′-biphenylene) ditetrazolium chloride (NT) reductase activity, whereas malonate, hydroquinone and ubiquinol-9 were ineffective. A differential activation of the flavoprotein by the oxidized and reduced forms of ubiquinone-9 was observed, the former stimulating the reduction of NT and the latter of phenazine methosulphate-2,6-dichlorophenolindophenol.4. Repeated washing of the activated mitochondrial samples with the sucrose homogenizing medium, partially reversed the activation by effectors other than succinate. Further washing of the activated preparations after a second preincubation with succinate reverted the enzyme activity to the basal level in the case of malonate, ATP and pyrophosphate but not that of hydroquinone and ubiquinol-9.5. Increase in the activity of hepatic mitochondrial succinate dehydrogenase, but not of succinate-NT reductase, known to occur in rats exposed to hypobaria was also observed in hypoxia indicating that it is an effect of lowered O₂ tension. The enzyme activity in these “partially activated” preparations was stable to washing with the sucrose homogenizing medium and could be fully activated to the same level as in the controls showing thereby the qualitative nature of the change. On washing these succinate-activated preparations further with the medium, the “hypobaric activation” was not reversed to the basal level, whereas the “hypoxic activation” was reversed. These results suggest that the effectors responsible for the activation of succinate dehydrogenase under hypobaric and hypoxic conditions are probably different; the former may be of the ubiquinol type and the latter of the malonate type.