scholarly journals Effect of environmental temperature on the kinetic properties of goldfish brain choline acetyltransferase

1972 ◽  
Vol 129 (5) ◽  
pp. 1013-1021 ◽  
Author(s):  
Catherine Hebb ◽  
T. C. Stephens ◽  
M. W. Smith
Keyword(s):  
Choline Acetyltransferase ◽  
Environmental Temperature ◽  
Incubation Temperature ◽  
Kinetic Properties ◽  
Stable Form ◽  
Acetyl Coa ◽  
Cold Adapted ◽  
Sequential Mechanism ◽  
Michaelis Constants ◽  
Environmental Temperatures

1. Michaelis constants of goldfish brain choline acetyltransferase were found to depend on the concentration of the second substrate present and on the temperature to which the fish had been adapted. 2. Primary plots constructed from results obtained with enzyme prepared from cold-adapted or warm-adapted fish indicated that synthesis of acetylcholine took place by a sequential mechanism. 3. The affinity of choline acetyltransferase for acetyl-CoA was about 100 times that for choline irrespective of whether the enzyme had been prepared from warm-adapted or cold-adapted fish. 4. The maximum rate at which choline acetyltransferase synthesized acetylcholine and the energy of activation for this synthesis remained independent of the previous environmental temperature of the fish. 5. The affinity of choline acetyltransferase for choline and acetyl-CoA showed a complex dependence on temperature. The affinity of the enzyme from cold-adapted fish for substrates increased as the incubation temperature was lowered, whereas that of the enzyme from warm-adapted fish first increased and then decreased. 6. The maximum affinity of choline acetyltransferase for both substrates, from both cold-adapted and warm-adapted fish, occurred at temperatures that corresponded approximately to the respective environmental temperatures of the fish. 7. These changes in enzyme affinity for substrates are not thought to be due to the presence of isoenzymes. Their adaptive significance is unknown, but it could be connected with the maintenance of the enzyme in a stable form.

scholarly journals The kinetic properties of human placental choline acetyltransferase

1971 ◽  
Vol 125 (3) ◽  
pp. 857-863 ◽  
Author(s):  
D. Morris ◽  
A. Maneckjee ◽  
Catherine Hebb
Keyword(s):  
Sodium Chloride ◽  
Choline Acetyltransferase ◽  
Product Inhibition ◽  
Rabbit Brain ◽  
Kinetic Properties ◽  
Forward Reaction ◽  
Acetyl Coa ◽  
No Inhibition ◽  
Ping Pong ◽  
Michaelis Constants

1. Michaelis constants for human placental choline acetyltransferase were shown to be dependent on the concentration of the second substrate present. The primary plots indicate a sequential rather than a Ping Pong mechanism and are of the same type with 300mm- and 500mm-sodium chloride. 2. Similar results have been obtained with rabbit brain choline acetyltransferase. 3. Product inhibition of the forward reaction has been studied. CoA inhibits competitively with respect to acetyl-CoA and non-competitively with respect to choline. Acetylcholine inhibits competitively with respect to choline and non-competitively with respect to acetyl-CoA. No inhibition is given by acetylcholine when the enzyme is saturated with choline. 4. It is concluded that human placental choline acetyltransferase has an ordered mechanism of the Theorell–Chance type.

scholarly journals The kinetic properties of citrate synthase from rat liver mitochondria

1969 ◽  
Vol 114 (3) ◽  
pp. 597-610 ◽  
Author(s):  
D. Shepherd ◽  
P. B. Garland
Keyword(s):  
Rat Liver ◽  
Citrate Synthase ◽  
Adenine Nucleotide ◽  
Sedimentation Coefficient ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Kinetic Properties ◽  
Acetyl Coa ◽  
Ph Optimum ◽  
Michaelis Constants

1. Citrate synthase (EC 4.1.3.7) was purified 750-fold from rat liver. 2. Measurements of the Michaelis constants for the substrates of citrate synthase gave values of 16μm for acetyl-CoA and 2μm for oxaloacetate. Each value is independent of the concentration of the other substrate. 3. The inhibition of citrate synthase by ATP, ADP and AMP is competitive with respect to acetyl-CoA. With respect to oxaloacetate the inhibition by AMP is competitive, but the inhibition by ADP and ATP is mixed, being partially competitive. 4. At low concentrations of both substrates the inhibition by ATP is sigmoidal and a Hill plot exhibits a slope of 2·5. 5. The pH optimum of the enzyme is 8·7, and is not significantly affected by ATP. 6. Mg2+ inhibits citrate synthase slightly, but relieves the inhibition caused by ATP in a complex manner. 7. At constant total adenine nucleotide concentration made up of various proportions of ATP, ADP and AMP, the activity of citrate synthase is governed by the concentration of the sum of the energy-rich phosphate bonds of ADP and ATP. 8. The sedimentation coefficient of the enzyme, as measured by activity sedimentation, is 6·3s, equivalent to molecular weight 95000.

scholarly journals Kinetic properties of the partially purified pyruvate dehydrogenase complex of ox brain

1973 ◽  
Vol 131 (1) ◽  
pp. 31-37 ◽  
Author(s):  
John P. Blass ◽  
Carole A. Lewis
Keyword(s):  
Pyruvate Dehydrogenase ◽  
Tricarboxylic Acid Cycle ◽  
Pyruvate Dehydrogenase Complex ◽  
Kinetic Properties ◽  
Acetyl Coa ◽  
Ph Optimum ◽  
Michaelis Constants ◽  
High Concentrations ◽  
Similar Preparation ◽  
Dehydrogenase Complex

The properties of a purified preparation of the pyruvate dehydrogenase complex from ox brain have been compared with those of a similar preparation from ox kidney. A broad pH optimum around 7.8, similar dependence on ionic strength, and independence of the nature of the buffer anions or cations characterized preparations from both tissues. Michaelis constants for the binding of pyruvate, thiamin pyrophosphate, NAD+ and CoA were also similar. Enzyme from both tissues was inhibited by NADH, by copper and other heavy metals, by high concentrations of tricarboxylic acid-cycle intermediates, and by preincubation with ATP. Acetyl-CoA itself did not appear to inhibit these preparations, although some commercial preparations of acetyl-CoA did contain an inhibitor. Although oxaloacetate and α-oxobutyrate were weak inhibitors, a number of other α-oxo acids including phenylpyruvate did not inhibit. The properties of the pyruvate dehydrogenase complex from brain and kidney appeared similar.

Dealing with hot rocky environments: Critical thermal maxima and locomotor performance in Leptodactylus lithonaetes (Anura: Leptodactylidae)

2019 ◽  
pp. 155-161 ◽  
Author(s):  
Ivan Beltran
Keyword(s):  
Environmental Temperature ◽  
Extreme Temperature ◽  
Effect Of Temperature ◽  
Maximum Speed ◽  
Locomotor Performance ◽  
Extreme Temperatures ◽  
Physiological Limits ◽  
Fitness Consequences ◽  
Thermal Maximum ◽  
Environmental Temperatures

Environmental temperature has fitness consequences on ectotherm development, ecology and behaviour. Amphibians are especially vulnerable because thermoregulation often trades with appropriate water balance. Although substantial research has evaluated the effect of temperature in amphibian locomotion and physiological limits, there is little information about amphibians living under extreme temperature conditions. Leptodactylus lithonaetes is a frog allegedly specialised to forage and breed on dark granitic outcrops and associated puddles, which reach environmental temperatures well above 40 ˚C. Adults can select thermally favourable microhabitats during the day while tadpoles are constrained to rock puddles and associated temperature fluctuations; we thus established microhabitat temperatures and tested whether the critical thermal maximum (CTmax) of L. lithonaetes is higher in tadpoles compared to adults. In addition, we evaluated the effect of water temperature on locomotor performance of tadpoles. Contrary to our expectations, puddle temperatures were comparable and even lower than those temperatures measured in the microhabitats used by adults in the daytime. Nonetheless, the CTmax was 42.3 ˚C for tadpoles and 39.7 ˚C for adults. Regarding locomotor performance, maximum speed and maximum distance travelled by tadpoles peaked around 34 ˚C, approximately 1 ˚C below the maximum puddle temperatures registered in the puddles. In conclusion, L. lithonaetes tadpoles have a higher CTmax compared to adults, suggesting a longer exposure to extreme temperatures that lead to maintain their physiological performance at high temperatures. We suggest that these conditions are adaptations to face the strong selection forces driven by this granitic habitat.

The effects of environmental temperature and method of feeding on the performance and carcass composition of bacon pigs

1967 ◽  
Vol 9 (2) ◽  
pp. 209-218 ◽  
Author(s):  
D. W. Holme ◽  
W. E. Coey
Keyword(s):  
Environmental Temperature ◽  
Average Daily Gain ◽  
Carcass Composition ◽  
Restricted Feeding ◽  
Feed Conversion ◽  
Feeding Method ◽  
Feeding Regimes ◽  
Higher Temperature ◽  
Ad Libitum ◽  
Environmental Temperatures

A trial designed to investigate the effects of two environmental temperatures, three feeding regimes and the interactions between them is described. A temperature of 72° F. was better than one of 54° F. for bacon pigs between 40 lb. and 200 lb. weight. The higher temperature resulted in faster growth, more efficient feed conversion and increased length of carcass. Other carcass characteristics were not significantly altered. Ad libitum feeding resulted in faster growth and fatter carcasses than restricted feeding, but did not have a significant effect on efficiency of feed conversion. When feed intake was restricted, feeding pigs once daily or twice daily resulted in similar performance and carcass composition.There was a significant interaction between environmental temperature and feeding method for average daily gain in that pigs fed ad libitum grew faster at the low temperature and pigs fed restricted amounts of feed grew faster at the high temperature. No other interaction reached significant levels.

Phosphoenolpyruvate Carboxylase of Thiobacillus thiooxidans. Kinetic and Metabolic Control Properties

1972 ◽  
Vol 50 (2) ◽  
pp. 158-165 ◽  
Author(s):  
R. L. Howden ◽  
H. Lees ◽  
Isamu Suzuki
Keyword(s):  
Enzyme Activity ◽  
Competitive Inhibitor ◽  
Acetyl Coa ◽  
Ph Optimum ◽  
Pep Carboxylase ◽  
Thiobacillus Thiooxidans ◽  
Powerful Activator ◽  
Michaelis Constants ◽  
Noncompetitive Inhibitor ◽  
Decreasing Ph

Phosphoenolpyruvate (PEP) carboxylase (orthophosphate:oxalacetate carboxy-lyase (phosphorylating), EC 4.1.1.31) was purified 19-fold from Thiobacillus thiooxidans. The level of enzyme activity was dependent on culture age. No enzyme activity could be obtained from frozen cells.The pH optimum of the enzyme was determined to be around 8.0. Apparent Michaelis constants were determined for the substrates:phosphoenolpyruvate (1.4, 1.5 mM), bicarbonate (0.4, 1.1 mM), and magnesium (1.1, 0.8 mM) at pH 7.0 and 8.0, respectively. Acetyl-CoA was found to be a powerful activator of this enzyme, with the degree of activation increasing with decreasing pH. The concentration of acetyl-CoA to obtain half-maximal activation, however, remained fairly constant and low, namely 1.2 and 1.0 μM at pH 7.0 and 8.0, respectively. L-Aspartate and L-malate were strong inhibitors of enzyme activity. In the presence of aspartate at pH 7.0 the double reciprocal activity plots for PEP became nonlinear, a characteristic of negative cooperativity. These plots became linear with the addition of acetyl-CoA with aspartate now acting as a noncompetitive inhibitor with respect to PEP. At pH 8.0, the same plots were linear with aspartate acting as a competitive inhibitor of PEP. All the other effectors of PEP carboxylase from Salmonella typhimurium and Escherichia coli were found to be ineffective towards the enzyme from T. thiooxidans.

scholarly journals Ligand binding on to maize (Zea mays) malate synthase: a structural study

1994 ◽  
Vol 303 (2) ◽  
pp. 413-421 ◽  
Author(s):  
S Beeckmans ◽  
A S Khan ◽  
L Kanarek ◽  
E Van Driessche
Keyword(s):  
Zea Mays ◽  
Ligand Binding ◽  
Conformational Change ◽  
Limited Proteolysis ◽  
Enzymic Activity ◽  
Malate Synthase ◽  
Acetyl Coa ◽  
Original Activity ◽  
Kinetic Measurements ◽  
Michaelis Constants

A kinetic and ligand binding study on maize (Zea mays) malate synthase is presented. It is concluded from kinetic measurements that the enzyme proceeds through a ternary-complex mechanism. Michaelis constants (Km,glyoxylate and Km,acetyl-CoA) were determined to be 104 microM and 20 microM respectively. C.d. measurements in the near u.v.-region indicate that a conformational change is induced in the enzyme by its substrate, glyoxylate. From these studies we are able to calculate the affinity for the substrate (Kd,glyoxylate) as 100 microM. A number of inhibitors apparently trigger the same conformational change in the enzyme, i.e. pyruvate, glycollate and fluoroacetate. Another series of inhibitors bearing more bulky groups and/or an extra carboxylic acid also induce a conformational change, which is, however, clearly different from the former one. Limited proteolysis with trypsin results in cleavage of malate synthase into two fragments of respectively 45 and 19 kDa. Even when no more intact malate synthase chains are present, the final enzymic activity still amounts to 30% of the original activity. If trypsinolysis is performed in the presence of acetyl-CoA, the cleavage reaction is appreciably slowed down. The dissociation constant for acetyl-CoA (Kd,acetyl-CoA) was calculated to be 14.8 microM when the glyoxylate subsite is fully occupied by pyruvate and 950 microM (= 50 x Km) when the second subsite is empty. It is concluded that malate synthase follows a compulsory-order mechanism, glyoxylate being the first-binding substrate. Glyoxylate triggers a conformational change in the enzyme and, as a consequence, the correctly shaped binding site for acetyl-CoA is created. Demetallization of malate synthase has no effect on the c.d. spectrum in the near u.v.-region. Moreover, glyoxylate induces the same spectral change in the absence of Mg2+ as in its presence. Nevertheless, malate synthase shows no activity in the absence of the cation. We conclude that Mg2+ is essential for catalysis, rather than for the structure of the enzyme's catalytic site.

scholarly journals Effects of 7°C environmental temperature acclimation during a 3-week training period

2020 ◽  
Vol 128 (4) ◽  
pp. 768-777
Author(s):  
Robert Shute ◽  
Katherine Marshall ◽  
Megan Opichka ◽  
Halee Schnitzler ◽  
Brent Ruby ◽  
...  
Keyword(s):  
Room Temperature ◽  
Environmental Temperature ◽  
Cellular Signaling ◽  
Temperature Acclimation ◽  
Peroxisome Proliferator ◽  
Peroxisome Proliferator Activated Receptor ◽  
Training Adaptations ◽  
Before And After ◽  
The Impact ◽  
Environmental Temperatures

Cold environmental temperatures during exercise and recovery alter the acute response to cellular signaling and training adaptations. Approximately 3 wk is required for cold temperature acclimation to occur. To determine the impact of cold environmental temperature on training adaptations, fitness measurements, and aerobic performance, two groups of 12 untrained male subjects completed 1 h of cycling in 16 temperature acclimation sessions in either a 7°C or 20°C environmental temperature. Fitness assessments before and after acclimation occurred at standard room temperature. Muscle biopsies were taken from the vastus lateralis muscle before and after training to assess molecular markers related to mitochondrial development. Peroxisome proliferator-activated receptor-γ coactivator 1α ( PGC-1α) mRNA was higher in 7°C than in 20°C in response to acute exercise before training ( P = 0.012) but not after training ( P = 0.813). PGC-1α mRNA was lower after training ( P < 0.001). BNIP3 was lower after training in the 7°C than in the 20°C group ( P = 0.017) but not before training ( P = 0.549). No other differences occurred between temperature groups in VEGF, ERRα, NRF1, NRF2, TFAM, PINK1, Parkin, or BNIP3L mRNAs ( P > 0.05). PGC-1α protein and mtDNA were not different before training, after training, or between temperatures ( P > 0.05). Cycling power increased during the daily training ( P < 0.001) but was not different between temperatures ( P = 0.169). V̇o2peak increased with training ( P < 0.001) but was not different between temperature groups ( P = 0.460). These data indicate that a 3-wk period of acclimation/training in cold environmental temperatures alters PGC-1α gene expression acutely but this difference is not manifested in a greater increase in V̇o2peak and is dissipated as acclimation takes place. NEW & NOTEWORTHY This study examines the adaptive response of cellular signaling during exercise in cold environmental temperatures. We demonstrate that peroxisome proliferator-activated receptor-γ coactivator 1α mRNA is different between cold and room temperature environments before training but after training this difference no longer exists. This initial difference in transcriptional response between temperatures does not lead to differences in performance measures or increases in protein or mitochondria.

LOW-TEMPERATURE ENVIRONMENTAL STRESS AND THE METABOLISM OF VITAMIN A IN THE RAT

1962 ◽  
Vol 40 (1) ◽  
pp. 491-499 ◽  
Author(s):  
W. E. J. Phillips
Keyword(s):  
Low Temperature ◽  
Vitamin A ◽  
Environmental Stress ◽  
Room Temperature ◽  
Environmental Temperature ◽  
Sprague Dawley ◽  
Vitamin A Metabolism ◽  
Environmental Temperatures

Carotene and vitamin A metabolism in the rat were studied at two environmental temperatures. The utilization of carotene is greater in animals maintained at a low environmental temperature (2°) than at room temperature (22°). This occurred within a period of 3 days. Both the hepatic storage and the rate of metabolism of orally administered vitamin A were unaffected by environmental temperature. The response of Wistar and Sprague–Dawley strains was similar.

scholarly journals Thermodynamic Analysis of Transcritical CO2 Ejector Expansion Refrigeration Cycle with Dedicated Mechanical Subcooling

Entropy ◽  
2019 ◽  
Vol 21 (9) ◽  
pp. 874
Author(s):  
Fu ◽  
Wang ◽  
Zheng ◽  
Yu ◽  
Liu ◽  
...  
Keyword(s):  
Thermodynamic Analysis ◽  
Environmental Temperature ◽  
Exergy Efficiency ◽  
Coefficient Of Performance ◽  
Refrigeration Cycle ◽  
Evaporation Temperature ◽  
Mass Ratios ◽  
Environmental Temperatures

: The new configuration of a transcritical CO2 ejector expansion refrigeration cycle combined with a dedicated mechanical subcooling cycle (EMS) is proposed. Three mass ratios of R32/R1234ze(Z) (0.4/0.6, 0.6/0.4, and 0.8/0.2) were selected as the refrigerants of the mechanical subcooling cycle (MS) to further explore the possibility of improving the EMS cycle’s performance. The thermodynamic performances of the new cycle were evaluated using energetic and exergetic methods and compared with those of the transcritical CO2 ejector expansion cycle integrated with a thermoelectric subcooling system (ETS). The results showed that the proposed cycle presents significant advantages over the ETS cycle in terms of the ejector performance and the system energetic and exergetic performances. Taking the EMS cycle using R32/R1234ze(Z) (0.6/0.4) as the MS refrigerant as an example, the improvements in the coefficient of performance and system exergy efficiency were able to reach up to 10.27% and 15.56%, respectively, at an environmental temperature of 35 C and evaporation temperature of −5 C. Additionally, the advantages of the EMS cycle were more pronounced at higher environmental temperatures.

Sign in / Sign up

Export Citation Format

Share Document