Rate Of ATP Consumption Research Articles

Protein Kinase A Does Not Alter Economy of Force Maintenance in Skinned Rat Cardiac Trabeculae

Recent mechanical, biochemical, and energetic experiments have suggested that catecholamines may increase the cycling rate of cross-bridges independent of changes inn intracellular calcium. An increased rate of cross-bridge cycling is expected to result in decreased economy of force maintenance. The present study tested this hypothesis directly by measuring the rate of ATP consumption in skinned cardiac trabeculae as a function of steady state force. Rat cardiac trabeculae were skinned with Triton X-100. Resting sarcomere length was measured by laser diffraction, and ATP consumption was assessed by an enzyme-coupled optical technique. Force-[Ca2+] relations were fit to a modified Hill equation. Force dependency of the rate of ATP consumption was analyzed by multiple linear regression analysis. beta-Adrenergic stimulation was mimicked by incubation of the skinned muscle preparation with the catalytic subunit of protein kinase A (PKA). Treatment with PKA (3 micrograms/mL, 40 minutes) induced a significant (65 +/- 23%, P = .01) increase in [Ca2+] required for half-maximal steady state force, whereas the steepness of the force-[Ca2+] relation was not affected. The rate of ATP consumption was linearly correlated with steady state force, regardless of PKA treatment status (P < .001). However, neither the slope nor the intercept was affected by PKA treatment. Hence, PKA treatment did not affect either the maximum rate of ATP consumption or the economy of force maintenance. These results suggest that beta-adrenergic stimulation does not alter the rate-limiting step of cross-bridge cycling during isometric contraction in myocardium.

A kinetic model for substrate and energy consumption of microbial growth under substrate-sufficient conditions.

The growth of heterotrophic microorganisms can be classified into substrate-limited and substrate-sufficient growth according to the relative availability of the substrate (carbon and energy source) and other nutrients. It is generally observed that the consumption rates of substrate and energy (ATP) are higher under substrate-sufficient conditions than under conditions of substrate limitation. The excess substrate and ATP consumption is often influenced by the residual concentration of substrate in a relatively wide range. To account for these effects, a kinetic model is proposed to describe substrate and ATP consumption rates of microbial growth under substrate-sufficient conditions. According to the model, the specific substrate consumption rate of a substrate-sufficient culture can be expressed as the sum of the substrate rate under substrate-limited conditions at the corresponding specific growth rate and an additional consumption rate due to excess substrate. The same kinetic form also applies to the specific ATP consumption rate and to the specific oxygen consumption rate of an aerobic culture, respectively. The linear equations for substrate and ATP consumption rates of Pirt and of Stouthammer and Bettenhousen can be used for substrate-limited growth. The excess of substrate and ATP consumption rates at carbon surplus can be described in a form similar to that of Michaelis-Menten kinetics. The proposed kinetic model has been verified with experimental data from three continuous cultures representing both anaerobic and aerobic microbial growth on substrates with low and high degrees of reductance. Using this model, the parameters maximum growth yield and maintenance requirement (both in terms of substrate and ATP) of a culture under different growth limitations can be better defined and quantified.(ABSTRACT TRUNCATED AT 250 WORDS)

Energy metabolism in muscle approaching maximal rates of oxygen utilization

Muscle is capable of operating over a wide range of metabolic rates. Most of the metabolic energy for work (ATP), and essentially all of the oxygen consumption of muscle, is due to mitochondrial oxidative phosphorylation. The rate of mitochondrial oxidative phosphorylation is determined by the rate of ATP consumption by the cells (demand) while the metabolic energy level at each metabolic rate is determined by the supply of oxidizable substrate and oxygen (supply). The maximal rate of metabolism is limited by several factors, including the supply of oxidizable substrates (mitochondrial dehydrogenases), of oxygen (blood supply and oxygen diffusion) and the respiratory enzyme capacity (tissue content of mitochondria).

Dependence of ATP consumption on cross-bridge phosphorylation in swine carotid smooth muscle.

1. Ca(2+)-dependent phosphorylation of the myosin regulatory light chain (MRLC) initiates cross-bridge cycling and contraction in smooth muscle. A four-state cross-bridge model, in which Ca(2+)-dependent phosphorylation is the only proposed regulatory mechanism, can predict the mechanical output of the swine carotid media. Our aims were to determine whether ATP consumption rates and the economy of force maintenance are regulated functions of MRLC phosphorylation as predicted by the model. 2. Steady-state force and oxygen consumption were measured in medial rings of swine carotid arteries activated with depolarizing solutions and agents capable of maintaining a wide range of steady-state myoplasmic Ca2+ and MRLC phosphorylation levels. 3. Suprabasal ATP consumption increased almost linearly with MRLC phosphorylation and exhibited a hyperbolic increase with active stress, as predicted. 4. The economy of stress maintenance fell with increases in suprabasal phosphorylation. 5. In absolute terms the energetic cost of covalent regulation by cross-bridge phosphorylation was small, although it may be a significant fraction of the ATP consumption associated with contraction.

Inhibitory mechanisms of antibody production by nitrogen oxides released from activated macrophages during the immune response: relationship to energy consumption.

We investigated the relationship between the sensitivity of mouse splenocytes in immune response to nitrogen oxides and energy consumption rate of the cells. Macrophage-like cells (Mm1) pretreated with IL-6 served as the source of the nitrogen oxides. The antibody production of both 2,4,6-trinitrophenyl-keyhole limpet haemocyanin-primed splenocytes and B cell hybridomas was markedly reduced; about 20-40% of splenocytes and B cell hybridomas were killed by co-culture with IL-6-treated Mm1. Cell viability and antibody production were completely restored by the addition of NG-monomethyl L-arginine to the culture medium. The cytotoxicity of the nitrogen oxides was correlated with the distance between effector and target cells. Under conditions of low cytotoxicity, antibody production by B cell hybridomas was suppressed by the nitrogen oxides, this suppression not being correlated with the reduction in cell growth. The sensitivity of the target cells differed in co-cultures of antigen-primed splenocytes and B cell hybridomas with IL-6-treated Mm1. The nitric oxide-sensitivity of the cells corresponded to their 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide reducing activity and ATP consumption rate. These findings suggest that nitrogen oxides act as regulatory molecules in immune response in three ways: cytostasis, reduction of cell growth and suppression of antibody synthesis.

What is special about smooth muscle? The significance of covalent crossbridge regulation.

Knowledge of the molecular mechanisms involved in chemomechanical transduction and the regulatory elements determining contraction and relaxation are largely derived from cross-striated muscle. However, it has become clear that vertebrate smooth muscles have features lacking in striated muscle. These derive from regulated crossbridge cycling rates observed as variable shortening velocities and ATP consumption rates. The special properties of smooth muscle are associated with differences in the physiological roles of muscle in the walls of hollow organs and reflect a common molecular motor governed by different regulatory mechanisms. A remarkably simple scheme involving Ca(2+)-dependent phosphorylation of crossbridges in smooth muscle can predict much of the mechanical and energetics behavior characterizing the muscle of hollow organs. Nevertheless, many unresolved issues are identified that are the focus of current research efforts.

Composition and Distribution of Adenylates in Soybean (Glycine max L.) Nodule Tissue.

Adenylates (ATP, ADP, and AMP) may play a central role in the regulation of the O2-limited C and N metabolism of soybean nodules. To be able to interpret measurements of adenylate levels in whole nodules and to appreciate the significance of observed changes in adenylates associated with changes in O2-limited metabolism, methods were developed for measuring in vivo levels of adenylate pools in the cortex, plant central zone, and bacteroid fractions of soybean (Glycine max L. Merr cv Maple Arrow x Bradyrhizobium japonicum strain USDA 16) nodules. Intact nodulated roots were either frozen in situ by flushing with prechilled Freon-113(-156[deg]C) or by rapidly (<1 s) uprooting plants and plunging them into liquid N2. The adenylate energy charge (AEC = [ATP + 0.5 x ADP]/[ATP + ADP + AMP]) of whole-nodule tissue (0.65 [plus or minus] 0.01, n = 4) was low compared to that of subtending roots (0.80 [plus or minus] 0.03, n = 4), a finding indicative of hypoxic metabolism in nodules. The cortex and central zone tissues were dissected apart in lyophilized nodules, and AEC values were 0.84 [plus or minus] 0.04 and 0.61 [plus or minus] 0.03, respectively. Although the total adenylate pool in the lyophilized nodules was only 41% of that measured in hydrated tissues, the AEC values were similar, and the lyophilized nodules were assumed to provide useful material for assessing adenylate distribution. The nodule cortex contained 4.4% of whole-nodule adenylates, with 95.6% being located in the central zone. Aqueous fractionation of bacteroids from the plant fraction of whole nodules and the use of marker enzymes or compounds to correct for recovery of bacteroids and cross-contamination of the bacteroid and plant fractions resulted in estimates that 36.2% of the total adenylate pool was in bacteroids, and 59.4% was in the plant fraction of the central zone. These are the first quantitative assessments of adenylate distribution in the plant and bacteroid fractions of legume nodules. These estimates were combined with theoretical calculations of rates of ATP consumption in the cortex (9.5 nmol g-1 fresh weight of nodule s-1), plant central zone (38 nmol g-1 fresh weight of nodule s-1), and bacteroids (62 nmol g-1 fresh weight of nodule s-1) of soybean nodules to estimate the time constants for turnover of the total adenylate pool and the ATP pool within each nodule fraction. The low values for time constant (1.6-5.8 s for total adenylate, 0.9-2.5 s for ATP only) in each fraction reflect the high metabolic activity of soybean nodules and provide a background for further studies of the role of adenylates in O2-limited nodule metabolism.

Bioenergetics and end‐product regulation of Clostridium thermosaccharolyticum in response to nutrient limitation

Fermentation of xylose by Clostridium thermosaccharolyticum was studied in batch and continuous culture in which the limiting nutrient was either xylose, phosphate, or ammonia. Transient results obtained in continuous cultures with batch grown inoculum and progressively higher feed substrate concentrations exhibited ethanol selectivities (moles ethanol/moles other products) in excess of 11. The hypothesis that this high ethanol selectivity was a general response to mineral nutrient limitation was tested but could not be supported. Growth and substrate consumption were related by the equation q(s)(1 - Y(x) (c))G(ATP) = (mu/Y(ATP) (max)) + m, with q(s) the specific rate of xylose consumption (moles xylose/hour . g cells), Y(x) (c) the carbon based cell yield (g cell carbon/g substrate carbon), G(ATP) the ATP gain (moles ATP produces/mol substrate catabolized), micro the specific growth rate (1/h), Y(ATP) (max) the ATP-based cell yield (g cells/mol ATP), and m the maintenance coefficient (moles ATP/hour . g cells). Y(ATP) (max) was found to be 11.6 g cells/mol ATP, and m 9.3 mol ATP/hour . g cells for growth on defined medium. Different responses to nutrient limitation were observed depending on the mode of cultivation. Batch and immobilized cell continuous cultures decreased G(ATP) by initiating production of the secondary metabolites, propanediol, and in some cases, D-lactate; in addition, batch cultures increased the fractional allocation of ATP to maintenance and/or wastage. Nitrogen-limited continuous free-cell cultures maintained a constant cell yield, whereas phosphate-limited continuous free-cell cultures did not. In the case of phosphate limitation, the decreased ATP demand associated with the lowered cell yield was accompanied by an increased rate of ATP consumption for maintenance and/or wastage. Neither nitrogen or phosphorus-limited continuous free-cell cultures exhibited an altered G(ATP) in response to mineral nutrient limitation, and neither produced secondary metabolites.

Effect of oxygen on steady-state product distribution inBacillus polymyxa fermentations

Bacillus polymyxa ferments glucose to 1-2,3 butanediol, acetoin, ethanol, acetic acid, lactic acid, and formic acid. This research investigates product formation as a function of oxygen availability. A predictive model that simulates product distribution at known oxygen transfer rates is developed on the hypothesis that, in an energy-limited environment, B. polymyxa utilizes glucose and oxygen in the most efficient manner. The efficiency of utilization of glucose and oxygen is measured in terms of the ATP yields of each oxidative pathway. The identity of the products constituting the profile at the given oxygen transfer rate is determined by comparing the ATP production and consumption rates. While the ATP generated is calculated from a knowledge of the oxygen transfer rate and ATP yields of the oxidative pathways, the ATP consumption is estimated by the Pirt expression in terms of growth- and nongrowth-associated components. The product formation rates are obtained by solving ATP and NAD balance equations. They equate the production and consumption rates of these intermediates and are derived from the pseudo-steady-state hypothesis. The model is applied to continuous culture systems that are both open and closed with respect to biomass. At a given oxygen transfer rate, dilution rate, and inlet glucose concentration, the model predicts steady-state concentrations of two dominant fermentation end products with the help of four parameters that can be determined from independent experiments. In contrast with earlier approaches, the experimental studies are carried out in continuous culture. Product profiles are obtained at various oxygen transfer rates, fer rates, inlet glucose concentrations, and dilution rates. The effect of pH on the relative distribution of products is also demonstrated. Results indicate that the model is fairly successful in predicting product profiles as a function of oxygen availability.

Adenosine 5'-triphosphate consumption by smooth muscle as predicted by the coupled four-state crossbridge model

We have proposed a four-state crossbridge model to explain contraction and the latch state in arterial smooth muscle. Ca(2+)-dependent crossbridge phosphorylation was the only postulated regulatory mechanism and the latchbridge (a dephosphorylated, attached crossbridge) was the only novel element in the model. In this study, we used the model to predict rates of ATP consumption by crossbridge phosphorylation (JPhos) and cycling (JCycle) during isometric and isotonic contractions in arterial smooth muscle; then we compared model predictions with experimental data. The model predicted that JPhos and JCycle were similar in magnitude in isometric contractions, and both increased almost linearly with myosin phosphorylation. The predicted relationship between isometric stress and ATP consumption was quasihyperbolic, but approximately linear when myosin phosphorylation was below 35%, in agreement with most of the available data. Muscle shortening increased the predicted values of JCycle up to 3.7-fold depending on shortening velocity and the level of myosin phosphorylation. The predicted maximum work output per ATP was 7.4-7.8 kJ/mol ATP and was relatively insensitive to changes in myosin phosphorylation. The predicted increase in JCycle with shortening was in agreement with available data, but the model prediction that work output per ATP was insensitive to changes in myosin phosphorylation was unexpected and remains to be tested in future experiments.

Rigor-Mortis Progress of Carp Acclimated to Different Water Temperatures.

Effects of the acclimation temperatures of carp between 5 and 30°C were examined on rigor-mortis progress and the postmortem changes of ATP and related compounds, glycogen, and lactate during storage at 0 and 10°C. The acclimation temperature gave large effects on both period required for reaching a maximal rigor and rigor strength. Rigor-mortis proceeded faster with increasing the difference between acclimation and storage temperatures. Rigor-mortis attained its maximum at 72h for 0 and 5°C-difference, while at 56, 32, and 24h for 10, 20, and 30°C-difference, respectively. For carp given 30°C temperature difference, maximal rigor index was almost 100%, while only below 60% for 0 and 5°C-difference. After transfer of carp to 30°C from 14°C, it took 4 weeks to change the rigor-mortis progress to a typical 30°C pattern. These data suggest that protein or lipid biosyntheses are involved in the changes of rigor-mortis patterns during temperature acclimation of carp. During the postmortem storage, changes of several compounds, i.e., ATP and glycogen de-crease and lactate accumulation, were faster for warm-acclimated carp than cold-acclimated one. These differences are considered to be derived from the differences of ATP consumption rates be-tween warm and cold-acclimated carp.

Intracellular calcium and high-energy phosphates in isolated cardiac myocytes

The relationship between intracellular calcium (Cai) and high-energy phosphates was studied in adult cardiac myocytes. Cai and high-energy phosphates were measured in the same population of cells. Cai, reported by the fluorescence of fura-2, was maintained at normal levels in the presence of increased transsarcolemmal calcium gradients, up to 5 mM extracellular calcium concentration. Cai was experimentally elevated by increasing calcium influx from the extracellular medium and/or by diminishing calcium efflux by Na-Ca exchange. Under these conditions, cells contracted and relengthened repetitively. The regulation of high-energy phosphates was challenged by increasing ATP utilization and by inhibiting ATP synthesis. Cai regulation was not affected by inhibition of glycolysis or NADH oxidation, so long as ATP concentration remained unchanged. High-energy phosphates were not depleted in beating cells with intact NADH oxidation, but inhibition of NADH oxidation caused a significant drop in phosphocreatine, demonstrating the increased rate of ATP consumption during beating. In beating cells, as in the working heart, ATP supply is increased to meet ATP demand, and steady-state ATP and phosphocreatine concentrations remain unchanged.

Balancing of energy‐consuming processes of rat hepatocytes

A method for the quantification of energy consuming processes described by Siems et al. for reticulocytes and by Müller et al. for ascites tumour cells was applied to balance the ATP-consumption of isolated rat hepatocytes. On the basis of decreased coupled respiration rates following the specific inhibition of energy-requiring reactions, the energy demands of protein turnover, nucleic acid synthesis, Na+/K(+)-ATPase and Ca2(+)-transport of hepatocytes in different incubation media were assessed. These processes together with urea synthesis account for about 60 per cent of the total energy consumption in a glucose and amino acid-enriched Eagle/Borsook medium. The metabolic flux rates of total ATP-consumption and ATP-consumption of single energy-requiring processes in hepatocytes are compared with those in reticulocytes and different tumour cell types.

Interrelationships (Between Glucose Metabolism, Energy State, and the Cytosolic Free Calcium Concentration in Cortical Synjaptosomes from the Guinea Pig

The stoichiometries of glycolysis and pyruvate oxidation were determined in cortical synaptosomes under varying rates of ATP consumption. Glycolysis was measured by using D-3-[3H]glucose as a marker and pyruvate oxidation by using D-3,4-[14C]glucose, which has to be metabolized to 1-[14C]pyruvate before being decarboxylated by the pyruvate dehydrogenase complex of intrasynaptosomal mitochondria. Cytosolic free Ca2+ concentration [( Ca2+]c) was determined in parallel and was manipulated by using EGTA in the incubation. The results show that in nonstimulated synaptosomes glycolysis and pyruvate oxidation are tightly coupled and stoichiometric. In the absence of Ca2+, when [Ca2+]c drops from 260 nM to 40 nM, glucose utilization increases, following the increase in energy demand, which has been shown to be due to elevated Na+ cycling. KCl depolarization, veratridine, and a mitochondrial uncoupler, carbonyl cyanide m-chlorophenylhydrazone, all stimulate glycolysis and pyruvate oxidation stoichiometrically, independently of the presence of external Ca2+. A rise in [Ca2+]c, therefore, is not required to regulate mitochondrial pyruvate metabolism. It is concluded that synaptosomes exhibit a high degree of respiratory control, that they rely on glucose oxidation for their energetics, and that stimulation of energy production can be achieved independently of changes in [Ca2+]c.

Laser radiation and motility patterns of human sperm.

Human sperm were exposed in vitro to laser radiation. An increase in progressive sperm motility was associated with a faster rate of sperm ATP consumption. Computer-assisted analysis of sperm motility confirmed the positive effect of laser irradiation on velocity and linearity of sperm.

31P NMR measurement of ATP synthesis rate in perfused intact rat hearts

Using 31P NMR and the saturation-transfer method, the unidirectional rate of ATP synthesis was measured in isolated, Langendorff-perfused, isovolumic rat hearts operating at a rate pressure product of 25.6 ± 2.5 (SE) × 10 3 mmHg · min −1 and consuming O 2; at a rate of 35 ± 2 μmol O 2 · min −1. (g dry wt) −1, at 37°C. This rate was 7.2 ± 0.9 μmol · s −1 · (g dry wt) −1 and was related to the rate of oxygen atom consumption by a ratio of 6.3 ± 0.9. These date show that in the intact heart the unidirectional rate of ATP synthesis exceeds the net rate of ATP synthesis and consumption by approximately a factor of 2.

Endogenous metabolism by sperm in response to altered cellular ATP requirements.

A quantitative description of the relative importance of endogenous metabolism to overall ATP production has not been established for mammalian cells. We report herein results of experiments using sperm (selected because of their simple metabolic potential and absence of biosynthetic pathways) and calorimetry (chosen because it serves as a general monitor of metabolism) to assess the importance of endogenous metabolism to total ATP synthesis under several incubation conditions. In experiments in which sperm were incubated at different temperatures (20 degrees C, 25 degrees C, 30 degrees C or 35 degrees C) and with different substrates (glucose, fructose, lactate, or beta-hydroxybutyrate), endogenous metabolism occurred at a constant rate regardless of the rate of ATP turnover in the cells or the nature of the exogenous substrate available to them. Sperm incubated at 35 degrees C with glycolyzable substrates synthesized more ATP (9 mumol ATP . h-1/10(8) cells) than did sperm incubated with the nonglycolyzable substrate, lactate (6.2 mumol ATP . h-1/10(8) cells). To investigate this substrate-related difference in the rate of ATP synthesis, the motility of sperm incubated at 35 degrees C with glucose or with lactate was determined. The velocities of the sperm incubated with either substrate were identical, indicating that the rates of ATP consumption for support of motility were identical. Most of the additional ATP synthesized by cells with glycolyzable substrates was consumed in the process of substrate cycling of the metabolic intermediates of glucose.

Energy requirements for stimulus-response coupling.

Platelets respond to numerous stimuli with many distinguishable responses. It has been well established that the receptor-mediated responses are absolutely dependent on the availability of ATP (1,2,3). This dependency may be due to ATP-requiring processes that 1) maintain the platelet in responsive state, 2) are involved in the stimulus-response coupling sequence and 3) drive the individual responses (Fig. 1). One way to indicate whether ATP consuming reactions participating in the stimulus-response coupling (signal processing) sequence would be to determine the rate of ATP consumption immediately after addition of agonist to platelets. The rapid (2–3 sec) fall in the metabolic adenylate charge in platelets upon ADP stimulation (4, 5) and increase in lactate-related H+ production after thrombin and ADP stimulation (6) suggest that ATP consumption is switched on immediately after agonist binding.KeywordsPhosphatidic AcidHuman PlateletPlatelet ResponseDense Granule SecretionWashed Rabbit PlateletThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Energy consumption in a cyclic phosphorylation/dephosphorylation cascade.

Cyclic phosphorylation/dephosphorylation cascade systems are responsible for regulating numerous metabolic pathways. The capacity of a cyclic cascade system to maintain a steady-state level of phosphorylation and, hence, a specific biological activity of a phosphorylatable protein is dependent upon a constant supply of metabolic energy (ATP). Quantification of the extent of ATP consumption in a cyclic cascade was examined experimentally with the model in vitro phosphorylation/dephosphorylation system described in detail in the previous paper (Shacter, E., Chock, P. B., and Stadtman, E. R. (1984) J. Biol. Chem. 259, 12252-12259). The results indicate that (a) when the concentrations of converter enzymes and interconvertible substrate are held constant and the fractional phosphorylation of the substrate is varied by changing the allosteric effector concentrations, the rate of ATP consumption in the monocyclic cascade is directly proportional to the steady-state level of phosphorylation being maintained. (b) Attainment of a particular steady-state level of phosphorylation is determined by the net ratio of the protein kinase and phosphatase activities and is independent of the absolute concentrations of these enzymes. (c) Whereas the time required to reach a given steady state is inversely proportional to the converter enzyme concentrations, the amount of ATP consumed in maintaining that steady state is directly proportional to the kinase and phosphatase concentrations. In addition, a theoretical analysis based upon experimentally determined parameters for two in vivo cyclic cascade systems (pyruvate kinase and glycogen phosphorylase) revealed that under normal conditions, cyclic phosphorylation/dephosphorylation cascades consume only a small proportion (less than 0.02%) of the total cellular energy flux.

Energy production during anoxia and recovery in the adductor muscle of the file shell, Lima hians

1. 1. During the initial period of anoxia (0–2.5 hr) 60% of the stored phosphagen of the adductor muscle was depleted, the levels of alanine increased and that of aspartate decreased. In the later period of anoxia (2.5–15 hr) there was a loss of 45% of the ATP content and the energy charge value dropped to 0.64. Lactate and octopine production were not observed. Formation of succinate took place during the whole anoxic period. 2. 2. During recovery the high energy status (phosphagen, ATP) was reached again after 1 hr as was the control level of succinate, whereas the concentration of alanine and aspartate were back to control values between 4 and 8 hr of recovery. 3. 3. The ATP consumption rate was greatly reduced in the later anoxic period. Glycolysis (first alanine, later succinate production) always contributed about 50%, arginine phosphate 47 and 27% and ATP 2 and 19% in the initial and later period, respectively.