The role of malate in hormone-induced enhancement of mitochondrial respiration

Shortly after the injection of glucagon, epinephrine, norepinephrine, vasopressin, or angiotensin II into fasted rats, mitochondria isolated from their livers contained elevated concentrations of malate and oxidized citrate, alpha-ketoglutarate, and, in some cases, succinate more rapidly than mitochondria from fasted, control rats. The administration of tryptophan, lactate, or ethanol and refeeding of rats fasted 24 h result in similar elevations of mitochondrial malate concentration and oxidation of added substrates. Treatments that resulted in elevated mitochondrial malate resulted also in increased uptake of added citrate, alpha-ketoglutarate, pyruvate, and, in some cases, succinate. It is postulated that the well-documented effect of gluconeogenic hormones on mitochondrial oxidation of carboxylic substrates may be mediated by malate which not only yields oxalacetate to support the tricarboxylic acid cycle but also facilitates the transport of added substrates, and which is regenerated in the tricarboxylic acid cycle.     

The control of mitochondrial respiration in yeast: A possible role of the outer mitochondrial membrane

Mitochondrial respiration in yeast (S. cerevisiae) is regulated by the level of glucose in the medium. Glucose is known to inhibit respiration by repressing key enzymes in the respiratory chain. We present evidence that the early events in this inhibition include the closure of VDAC channels, the primary pathway for metabolite flow across the outer membrane. Aluminum hydroxide is known to inhibit the closure of VDAC. Addition of aluminum acetylacetonate to yeast cells, which should elevate the aluminum hydroxide concentrations in the cytoplasm, caused the inhibition of cell respiration by glucose to be delayed for up to 100 min. No significant effect of aluminum was observed in cells grown on glycerol. Yeast cells lacking the VDAC gene were also unresponsive to the addition of aluminum salt in the presence of glucose. Therefore, the closure of VDAC channels may be an early step in the inhibition of the respiration of yeast by glucose.     

The role of malate in regulating the rate of mitochondrial respiration in vitro

Depletion of endogenous malate by preincubation of mitochondria at 30 degrees C in substrate-free media sharply decreases the rate of citrate oxidation and inhibits mitochondrial respiration in the presence of pyruvate and alpha-ketoglutarate. Addition of catalytic amounts of endogenous malate and its production via succinate oxidation promote rapid oxidation of citrate and pyruvate in the mitochondria and abolishes the lag period with alpha-ketoglutarate Malate increases the rate of membrane potential generation after addition of citrate, pyruvate or alpha-ketoglutarate to mitochondrial suspensions. Studies with controlled malate concentrations revealed that the changes in malate concentrations observed in the mitochondria in the presence of gluconeogenesis-inducing hormones may be due to the influence of these hormones on mitochondrial oxidation.     

The role of respiration during adaptation of the freshwater cyanobacterium Synechococcus 6311 to salinity

Growth of the freshwater cyanobacterium Synechococcus 6311 under saline conditions stimulated respiration tenfold during the first 24 h, while growth and photosynthesis were inhibited. The elevated respiration rate was seen under both light and dark conditions, was uncoupler and cyanide sensitive, and did not decrease upon salt removal. Membrane preparations from salt-grown cells exhibited a tenfold increase in cytochrome oxidase activity, while electron transfer rates from NADPH to cytochrome c only increased threefold. Cytochrome oxidase activities were correlated with levels of EPR detectable Cu2+ in the salt and control membranes. Sodium-driven proton (antiproter) gradients in salt-grown cells were sensitive to cyanide but not dicyclohexylcarbodiimide, indicating the direct role of respiratory electron transport in maintaining low intracellular sodium levels.     

The role of a protein factor in the effect of alpha-tocopherol on mitochondrial respiration

It is shown that alpha-tocopherol in vitro stimulates respiration of the liver mitochondria in E-hypovitaminosis rats only in the presence of the specific protein factor isolated from the liver cytosol. The action of alpha-tocopherol on mitochondria in the presence of NAD and a protein factor is not accompanied by an increase in the NADH level, that evidences for the absence of the direct redox interaction between NAD and tocopherol.     

Nitric oxide inhibition of mitochondrial respiration and its role in cell death

Nitric oxide (NO) or its derivatives (reactive nitrogen species, RNS) inhibit mitochondrial respiration in two different ways: (i) an acute, potent, and reversible inhibition of cytochrome oxidase by NO in competition with oxygen; and, (ii) irreversible inhibition of multiple sites by RNS. NO inhibition of respiration may impinge on cell death in several ways. Inhibition of respiration can cause necrosis and inhibit apoptosis due to ATP depletion, if glycolysis is also inhibited or is insufficient to compensate. Inhibition of neuronal respiration can result in excitotoxic death of neurons due to induced release of glutamate and activation of NMDA-type glutamate receptors. Inhibition of respiration may cause apoptosis in some cells, while inhibiting apoptosis in other cells, by mechanisms that are not clear. However, NO can induce (and inhibit) cell death by a variety of mechanisms unrelated to respiratory inhibition.     

Wheat mitochondrial proteomes provide new links between antioxidant defense and plant salinity tolerance

The mitochondrial proteome and differences associated with salt tolerance have been investigated in Australian commercial varieties of wheat. Mitochondria isolated from shoots were used to generate a wheat mitochondrial reference map; 68 unique wheat mitochondrial proteins were identified from 192 gel spots using 2D PAGE and LC-MS/MS. This analysis also provided MS/MS spectra for 199 proteotypic peptides as a foundation for the development of targeted proteomics to study the respiratory apparatus in wheat. Using this reference map and 2D DIGE, we have found quantitative differences in the shoot mitochondrial proteomes of v. Wyalkatchem and v. Janz, two commercially important wheat varieties that are known from a range of experiments to differ in salinity tolerance. These proteins included Mn-superoxide dismutase (Mn-SOD), cysteine synthase, nucleotide diphosphate kinase, and the voltage dependent anion channel (VDAC). Antibodies to the mitochondrial alternative oxidase (AOX), previously linked to reduced ROS formation from the electron transport chain and salt tolerance in Arabidopsis, also showed a commensurate higher abundance in v. Wyakatchem in both control and salt-treated conditions. Together, the data presented here suggest that differences in mitochondrial ROS defense pathways in the mitochondrial proteomes of key Australian wheat varieties correlate with whole-plant salinity tolerance.     

Mechanisms of salinity tolerance in plants

The mechanisms of salt stress response and tolerance have eluded definition despite reasonable success in defining their physiological manifestations. In this review, we consider the integrated salt metabolism of plants, essentially as a problem in meganutrient physiology. Two critical aspects of cellular and organismal metabolism are given particular attention—those involved in the control and integration of Na⁺ acquisition and allocation in plants and those involved in readjustment of other aspects of metabolism, especially those involving carbon as a resource.     

The role of salinity tolerance and competition in the distribution of an endangered desert salt marsh endemic

Rare plants are often associated with distinctive soil types, and understanding why endemic species occur in unique environments is fundamental for their management. At Ash Meadows National Wildlife Refuge in southern Nevada, USA, we evaluated whether the limited distribution of endangered Amargosa niterwort (Nitrophila mohavensis) is explained by this species’ tolerance of saline soils on salt-encrusted mud flats compared with the broadly distributed desert saltgrass (Distichlis spicata var. stricta). We simultaneously explored whether niterwort distribution is restricted from expanding due to interspecific competition with saltgrass. Surface soils collected throughout niterwort’s range were unexpectedly less saline with lower extractable Na, seasonal electroconductivity, and Na absorption ratio, and higher soil moisture than in adjacent saltgrass or mixed shrub habitats. Comparison of niterwort and saltgrass growth along an experimental salinity gradient in a greenhouse demonstrated lower growth of niterwort at all but the highest NaCl concentrations. Although growth of niterwort ramets was similar when transplanted into both habitats at the refuge below Crystal Reservoir, niterwort reproductive effort was considerably higher in saltgrass compared to its own habitat, implying reallocation of resources to sexual reproduction to maximize fitness when the probability of ramet mortality increases with greater salinity stress. Saltgrass was not a demonstrated direct competitor of niterwort; however, this species is known to increase soil salinity by exuding salt ions and through litterfall. Niterwort conservation will benefit from protecting hydrological processes that reduce salinity stress and preventing saltgrass colonization into niterwort habitat.     

The site of inhibition of dibromothymoquinone in mitochondrial respiration

Dibromothymoquinone (2,5-dibromo, 6-isopropyl, 3-methyl benzoquinone, DBMIB) is a quinone analogue recently introduced as a specific inhibitor of chloroplast photosynthesis at the level of plastoquinone. In beef heart mitochondria DBMIB inhibits the oxidation of both succinate and NAD linked substrates; the apparent K_I is 6 μM for βhydroxybutyrate oxidation and 61μM for succinate oxidation respectively. In sonic fragments NADH oxidation is also inhibited; however, the rotenone block of respiration can be partially bypassed by the autooxidation of reduced DBMIB. Under the same conditions succinoxidase of ETP is inhibited, as in intact mitochondria; autoxidation of DBMIB reduced by succinate can however be obtained in presence of detergents. Hexahydrocoenzyme Q₄ reverses the DBMIB inhibition of succinate in sonic fragments. The site of inhibition by DBMIB is the oxygen side of CoQ, since DBMIB can function as electron acceptor in the NADH-CoQ assay for site I energization in submitochondrial particles, studied by measuring the quenching of atebrin fluorescence.     

New insights into the role of Arabidopsis RABA1 GTPases in salinity stress tolerance

RAB11 GTPases, widely conserved members of RAB small GTPases, have evolved in a unique way in plants; plant RAB11 has notable diversity compared with animals and yeast. Recently, we have shown that members of RABA1, a subgroup in Arabidopsis RAB11 group, are required for salinity stress tolerance. To obtain a clue to understand its underlying mechanism, here we investigate whether RABA1 regulates sodium transport across the plasma membrane and accumulation in the vacuole. The results indicate that the raba1 quadruple mutant is not defective in the import and intracellular distribution of sodium, implying that RABA1 members are involved in a more indirect way in the responses to salinity stress.     

Recent developments in understanding salinity tolerance

Salt stress imposes a major environmental threat to agriculture and its adverse impacts are getting more serious problem in regions where saline water is used for irrigation. Therefore, the efforts to increase salt tolerance of crop plants bear remarkable importance to supply sustainable agriculture on marginal lands and could potentially improve crop yield overall. Acclimation of plants to salinized conditions depend upon activation of cascades of molecular networks involved in stress sensing, signal transduction and the expression of specific stress-related genes and metabolites. Adaptational processes are elaborate and more than one gene might be expressed during the acclimation process. Isolation of Salt Overly Sensitive (SOS) genes by sos mutants shed us light on the relationship between ion homeostasis and salinity tolerance. The essential role of antioxidative system to maintain a balance between the overproduction of Reactive Oxygen Species (ROS) and their scavenging to keep them at signaling level for reinstating metabolic homeostasis has already been established. Compatible osmolytes synthesized to maintain equal water potential with the environment under salinity conditions implements another strategy to develop resistance against salinity. With the growing body of information about molecular markers, genomics and post-genomics and thus increasing understanding of signaling pathways and mechanisms that contributes to plant stress responses, significant breakthroughs have been emerged to figure out the mechanism and control of salinity tolerance at molecular level. Many transgenic works were carried out to produce transgenic plants to develop enhanced tolerance to salt stress. However, a few of them seem succeeded to be implemented in salt-affected marginal lands efficiently. This minireview focuses on the recent developments in salinity tolerance research aiming to contribute sustainable food production under salt stress in the face of a globally warming ecosystem.     

Regulation of mitochondrial respiration in senescence

The ADP-stimulated (State 3) respiration of myocardial mitochondria with glutamate-malate, glutamate-pyruvate, palmitylcarnitine and β-hydroxybutyrate as substrates declined in rats after the age of 20 months. There was no significant decline in pyruvate-malate, α-oxoglutarate, palmityl-CoA, succinate and ascorbate cytochrome c oxidation. Skeletal muscle mitochondria from senescent animals showed a similar decline in glutamate-malate oxidation but not in palmityl-CoA, palmitylcarnitine, succinate and ascorbate-cytochrome c oxidation. The controlled oxidation with ADP-limiting (State 4) and the ADP/O ratio were not affected. The results indicate an alteration in the subtle regulatory capacity for mitochondrial oxidation in senescent rats. It is suggested that the alteration may be in certain anion transport and associated functions across the mitochondrial membrane or dehydrogenase activity.     

Control of mitochondrial respiration in muscle

Summary Control of mitochondrial respiration depends on ADP availability to the F₁ATPase. An electrochemical gradient of ADP and ATP across the mitochondrial inner membrane is maintained by the adenine nucleotide translocase which provides ADP to the matrix for ATP synthesis and ATP for energy-dependent processes in the cytosol. Mitochondrial respiration is responsive to the cytosolic phosphorylation potential, ATP/ADP · P_i which is in apparent equilibrium with the first two sites in the electron transport chain. Conventional measures of free adenine nucleotides is a confounding issue in determining cytosolic and mitochondrial phosphorylation potentials. The advent of phosphorus-31 nuclear magnetic resonance (P-31 NMR) allows the determination of intracellular free concentrations of ATP, creatine-P and Pi in perfused muscle in situ. In the glucose-perfused heart, there is an absence of correlation between the cytosolic phosphorylation potential as determined by P-31 NMR and cardiac oxygen consumption over a range of work loads. These data suggest that contractile work leads to increased generation of mitochondrial NADH so that ATP production keeps pace with myosin ATPase activity. The mechanism of increased ATP synthesis is referred to as stimulusre-sponse-metabolism coupling. In muscle, increased contractility is a result of interventions which increase cytosolic free Ca²⁺ concentrations. The Ca^2- signal thus generated increases glycogen breakdown and myosin ATPase in the cytosol. This signal is concomitantly transmitted to the mitochondria which respond to small increases in matrix Ca²⁺ by activation of Ca²⁺-sensitive dehydrogenases. The Ca²⁺-activated dehydrogenase activities are key rate-controlling enzymes in tricarboxylic acid cycle flux, and their activation by Ca^2- leads to increased pyridine nucleotide reduction and oxidative phosphorylation. These observations which have been consistent in preparations both in vitro and in situ do not obviate a role for ADP control of muscle respiration, but do explain, in part, the lack of dramatic fluctuations in the cytosolic phosphorylation potential over a large range of contractile activities.     

Control of plant mitochondrial respiration

Plant mitochondria are characterised by the presence of both phosphorylating (cytochrome) and non-phosphorylating (alternative) respiratory pathways, the relative activities of which directly affect the efficiency of mitochondrial energy conservation. Different approaches to study the regulation of the partitioning of reducing equivalents between these routes are critically reviewed. Furthermore, an updated view is provided regarding the understanding of plant mitochondrial respiration in terms of metabolic control. We emphasise the extent to which kinetic modelling and 'top-down' metabolic control analysis improve the insight in phenomena related to plant mitochondrial respiration. This is illustrated with an example regarding the affinity of the plant alternative oxidase for oxygen.