Maximal Activities Of Key Enzymes Research Articles

Citrulline enhances myofibrillar constituents expression of skeletal muscle and induces a switch in muscle energy metabolism in malnourished aged rats

Citrulline (Cit) actions on muscle metabolism remain unclear. Those latter were investigated using a proteomic approach on Tibialis muscles from male Sprague-Dawley rats. At 23 months of age, rats were either fed ad libitum (AL group) or subjected to dietary restriction for 12 weeks. At the end of the restriction period, one group of rats was euthanized (R group) and two groups were refed for one week with a standard diet supplemented with nonessential amino acids group or Cit (CIT group). Results of the proteomic approach were validated using targeted Western blot analysis and assessment of gene expression of the related genes. Maximal activities of the key enzymes involved in mitochondrial functioning were also determined. Cit supplementation results in a significant increase in the protein expression of the main myofibrillar constituents and of a few enzymes involved in glycogenolysis and glycolysis (CIT vs. AL and R, p < 0.05). Conversely, the expression of oxidative enzymes from Krebs cycle and mitochondrial respiratory chain was significantly decreased (CIT vs. AL, p < 0.05). However, maximal activities of key enzymes of mitochondrial metabolism were not significantly affected, except for complex 1 which presented an increased activity (CIT vs. AL and R, p < 0.05). In conclusion, Cit supplementation increases expression of the main myofibrillar proteins and seems to induce a switch in muscle energy metabolism, from aerobia toward anaerobia.

Metabolic rate does not scale with body mass in cultured mammalian cells

The allometric scaling of metabolic rate with organism body mass can be partially accounted for by differences in cellular metabolic rates. For example, hepatocytes isolated from horses consume almost 10-fold less oxygen per unit time as mouse hepatocytes [Porter and Brand, Am J Physiol Regul Integr Comp Physiol 269: R226-R228, 1995]. This could reflect a genetically programmed, species-specific, intrinsic metabolic rate set point, or simply the adaptation of individual cells to their particular in situ environment (i.e., within the organism). We studied cultured cell lines derived from 10 mammalian species with donor body masses ranging from 5 to 600,000 g to determine whether cells propagated in an identical environment (media) exhibited metabolic rate scaling. Neither metabolic rate nor the maximal activities of key enzymes of oxidative or anaerobic metabolism scaled significantly with donor body mass in cultured cells, indicating the absence of intrinsic, species-specific, cellular metabolic rate set points. Furthermore, we suggest that changes in the metabolic rates of isolated cells probably occur within 24 h and involve a reduction of cellular metabolism toward values observed in lower metabolic rate organisms. The rate of oxygen delivery has been proposed to limit cellular metabolic rates in larger organisms. To examine the effect of oxygen on steady-state cellular respiration rates, we grew cells under a variety of physiologically relevant oxygen regimens. Long-term exposure to higher medium oxygen levels increased respiration rates of all cells, consistent with the hypothesis that higher rates of oxygen delivery in smaller mammals might increase cellular metabolic rates.

The metabolic organization of a primitive air‐breathing fish, the Florida gar (lepisosteus platyrhincus)

The metabolic organization of the air-breathing Florida gar, Lepisosteus platyrhincus, was assessed by measuring the maximal activities of key enzymes in several metabolic pathways in selected tissues, concentrations of plasma metabolites including nonesterified fatty acids (NEFA), free amino acids (FAA) and glucose as well as tissue FAA levels. In general, L. platyrhincus has an enhanced capacity for carbohydrate metabolism as indicated by elevated plasma glucose levels and high activities of gluconeogenic and glycolytic enzymes. Based upon these properties, glucose appears to function as the major fuel source in the Florida gar. The capacity for lipid metabolism in L. platyrhincus appears limited as plasma NEFA levels and the activities of enzymes involved in lipid oxidation are low relative to many other fish species. L. platyrhincus is capable of oxidizing both D- and L-beta-hydroxybutyrate, with tissue-specific preferences for each stereoisomer, yet the capacity for ketone body metabolism is low compared with other primitive fishes. Based on enzyme activities, the metabolism of the air-breathing organ more closely resembles that of the mammalian lung than a fish swim bladder. The Florida gar sits phylogenetically and metabolically in an intermediate position between the "primitive" elasmobranchs and the "advanced" teleosts. The apparently unique metabolic organization of the gar may have evolved in the context of a bimodal air-breathing environmental adaptation.

Metabolic reorganization and signal transduction during estivation in the spadefoot toad

The maximal activities of 28 enzymes, representing multiple pathways of intermediary metabolism, were quantified in the brain, liver and skeletal muscle of spadefoot toads Scaphiopus couchii, comparing control toads with animals that had estivated for 2 months. Estivation-induced changes in brain enzyme activities were consistent with suppressed glycolysis and increased ketone body and amino acid catabolism. In liver, estivation resulted in reduced activities of eight enzymes representing carbohydrate, amino acid, ketone body and phosphagen metabolism, but the maximal activity of malic enzyme increased by 2.4-fold. Estivation led to a large-scale reorganization of skeletal muscle affecting most of the enzymes analyzed. Activities of enzymes of carbohydrate catabolism were generally elevated except for glycogen phosphorylase and hexokinase, whereas those of enzymes of fatty acid synthesis and ketone body metabolism were reduced. Increased glutamate dehydrogenase activities in both brain and muscle, as well as activities of other amino-acid-catabolizing enzymes in muscle, correlated with specific changes in the free amino acids pools in those tissues (reduced glutamine activity, increased glutamate, alanine and valine activities) that appear to be related to protein catabolism, for the purposes of elevating urea levels. The effects of estivation on signal transduction systems were also assessed. Total activities of protein kinases A and C (PKA and PKC) were largely unaltered in toad tissues during estivation (except for a 57% reduction in liver total PKC), but in seven organs there were strong reductions in the percentage of PKA present as the active catalytic subunit in estivating animals, and three contained a much lower percentage of membrane-bound active PKC during estivation. Activities of protein phosphatase types 1, 2A, 2B, and 2C were also frequently reduced during estivation. Overall, these results suggest that anuran estivation involves metabolic reorganization, including changing the maximal activities of key enzymes of intermediary metabolism as well as depressing the metabolic rate by suppressing signal transducing enzymes.

Maximal activities of enzymes of intermediary metabolism in the estivating terrestrial snail Cepaea nemoralis

The effects of estivation on maximal activities of key enzymes of aerobic, carbohydrate, lipid, ketone body and amino acid metabolism were studied in tissues of the terrestrial snail Cepaea nemoralis. With the exception of a 40% decrease in citrate synthase activity in hepatopancreas, enzyme activities were unaltered in all tissues following 6 weeks of estivation. Activities of enzymes of lipid, ketone body and carbohydrate metabolism were reduced in the kidney, when expressed on a per g wet tissue weight basis. However, estivation resulted in a near-doubling of kidney weight, though protein concentration of this tissue was decreased by 27%. When enzyme activities were expressed as units per mg −1 protein, no changes were observed with estivation in kidney. It was suggested that the changes in kidney weight are due to the storage of metabolic by-products, perhaps uric acid. No change in weight or protein content of heart or hepatopancreas were observed. These results suggest that no universal down-regulation of enzyme activities occurs during long-term estivation.

Effects of hypothyroidism on glucose and glutamine metabolism by the gut of the rat.

1. The metabolism of glucose and glutamine was studied in the small intestine and the colon of rats after 4-5 weeks of hypothyroidism. 2. Hypothyroidism resulted in increases in the plasma concentrations of ketone bodies (P less than 0.05), cholesterol (P less than 0.001) and urea (P less than 0.001), but decreases in the plasma concentrations of free fatty acids (P less than 0.05) and triacylglycerol (P less than 0.001). These changes were associated with decreases in the plasma concentrations of total tri-iodothyronine, free tri-iodothyronine, total thyroxine and free thyroxine. 3. Hypothyroidism decreased both the DNA content (by 30.5%) and the protein content (by 23.6%) of intestinal mucosa, with the protein/DNA ratio remaining unchanged. The villi in the jejunum were shorter (P less than 0.05) and the crypt depth was decreased by about 26.5% in hypothyroid rats. 4. Portal-drained visceral blood flow showed no marked change in response to hypothyroidism, but was accompanied by decreased rates of extraction of glucose, lactate and glutamine and release of glutamate, alanine and ammonia. 5. Enterocytes and colonocytes isolated from hypothyroid rats showed decreased rates of utilization and metabolism of glucose and glutamine. 6. The maximal activities of hexokinase (EC 2.7.1.1), 6-phosphofructokinase (EC 2.7.1.11), pyruvate kinase (EC 2.7.1.40), citrate synthase (EC 4.1.3.28), oxoglutarate dehydrogenase (EC 1.2.4.2) and phosphate-dependent glutaminase (EC 3.5.1.2) were decreased in intestinal mucosal scrapings from hypothyroid rats. Similar decreases were obtained in colonic mucosal scrapings (except for citrate synthase and oxoglutarate dehydrogenase) from hypothyroid rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Impact of an Acute Temperature Change on Performance and Metabolism of Pickerel (Esox niger) and Eel (Anguilla rostrata) Hearts

Mechanical performance, oxygen consumption (Ṁo2), and the maximal activities of key enzymes of energy metabolism were assessed in chain pickerel (Esox niger) and American eel (Anguilla rostrata) hearts over a temperature range normally experienced by these fish. Ventricle strips from pickerel could be paced up to 48 contractions min−1 at both 5° and 15°C. Eel ventricle strips responded at 48 contractions min−1 at 15° C but at 5° C could not be paced faster than 18 contractions min−1. At low temperatures, time to 50% relaxation was especially prolonged in eel ventricle strips, and increases in extracellular calcium resulted in large increases in resting tension. These findings suggest that calcium extrusion from the myocyte is impaired. Maximum output of perfused, isolated pickerel hearts occurred at frequencies of 30 and 36 contractions min−1 at 5° and 15°C. Values for eel hearts were 18 and 30 contractions min−1. Pickerel hearts had the same power output at both temperatures. Power output of eel hearts was lower at 5° than 15°C. Oxygen consumption of perfused hearts displayed a Q10 of about 1.9 for both pickerel and eel. Maximal in vitro activities of hexokinase, carnitine acyl-CoA transferase, citrate synthase, and ATPase were determined at 5° and 15° C. Comparison of Ṁo2 values with enzyme activities reveals that the catalytic potential to provide ATP is in excess of resting energy demand at both temperatures. The impairment of eel heart performance at low temperature cannot be attributed to an inability to metabolize glucose or fatty acids. The findings with isolated preparations are consistent With maintenance of activity by pickerel and presumptive in situ cardiac function over a wide temperature range and with the lack of activity displayed by eel at low temperature in association with known reduced cardiac performance.

Maximal activities of key enzymes of glutaminolysis, glycolysis, Krebs cycle and pentose-phosphate pathway of several tissues in mature and aged rats

1. 1. The maximum activities of some key enzymes, which provide a quantitative indices of flux through several important pathways have been measured in brain, liver, muscle, white and brown adipose tissue and lymphocytes of mature and aged rats. 2. 2. The results were expressed as pmol/min per g fresh weight and nmol/min per mg protein. 3. 3. On the both basis, as compared to mature rats, hexokinase activity is decreased in brown adipose tissue and increased in soleus muscle. 4. 4. Glucose-6-phosphate dehydrogenase activity is decreased in most tissues and increased in brain. 5. 5. Citrate synthase activity, which provides a qualitative index of the Krebs cycle, is decreased in white adipose tissues and lymphocytes. 6. 6. Glutaminase activity is decreased in brain, white and brown adipose tissues but is increased in lymphocytes.

Effect of glucocorticoid treatment on glucose and glutamine metabolism by the small intestine of the rat.

1. The effect of dexamethasone (30 micrograms day-1 100 g-1 body wt.) on the metabolism of glucose and glutamine was studied in the small intestine of rats after 9 days of treatment. 2. Dexamethasone treatment resulted in negative nitrogen balance (P less than 0.001), and produced increases in the concentrations of plasma glucose (22%, P less than 0.05), alanine (32%, P less than 0.001) and insulin (127%, P less than 0.001), but a decrease in the plasma concentration of glutamine (20%, P less than 0.05). 3. Portal-drained visceral blood flow increased by approximately 22% (P less than 0.001) in dexamethasone-treated rats, and was accompanied by a decrease in the arterio-venous concentration difference of glucose (43%, P less than 0.001) and an increase in that of lactate (22%, P less than 0.05), glutamine (35%, P less than 0.01), glutamate (33%, P less than 0.01) and alanine (21%, P less than 0.05). 4. Enterocytes isolated from dexamethasone-treated rats showed decreased and increased rates of glucose and glutamine utilization, respectively. 5. The maximal activities of hexokinase, 6-phosphofructokinase, citrate synthase and oxoglutarate dehydrogenase were decreased (30-64%, P less than 0.001) in intestinal mucosal scrapings of dexamethasone-treated rats, whereas the activity of glutaminase was increased (35%, P less than 0.001). 6. It is concluded that glucocorticoid administration decreases the rate of glucose utilization but increases that of glutamine (both in vivo and in vitro) by the epithelial cells of the small intestine. This may be caused by changes in the maximal activities of key enzymes in the pathways of glucose and glutamine metabolism in these cells.

Myoglobin content and the activities of enzymes of energy metabolism in red and white fish hearts

The myoglobin content of representative red and white coloured fish hearts was quantitated. It was confirmed that the macroscopic difference in appcarance is due to the presence or absence of myoglobin. Thereafter, the cytochrome c content as well as the maximal activities of key enzymes of energy metabolism were assessed in myoglobin-rich sea raven (Hemitripterus americanus) and myoglobin-poor ocean pout (Macrozoarces americanus) hearts. Both species are sluggish benthic dwellers that occur in similar habitats in the North Atlantic Ocean. The activities of enzymes associated with aerobic metabolism were similar in sea raven and ocean pout hearts, but far in excess of activities observed from white skeletal muscle. The two hearts also displayed comparable activities of enzymes associated with anaerobic energy metabolism. It therefore appears that the capacity to produce reducing equivalents for the electron transport system is similar in two selected fish hearts despite great differences in myoglobin content.