Proton pumping complex I increases growth yield in Candida utilis.

In living cells, growth is the result of coupling between substrate catabolism and multiple metabolic processes that take place during net biomass formation and cellular maintenance processes. A crucial parameter for growth evaluation is its yield, i.e.

The role of mitochondrial biogenesis and ROS in the control of energy supply in proliferating cells.

In yeast, there is a constant growth yield during proliferation on non-fermentable substrate where the ATP generated originates from oxidative phosphorylation. This constant growth yield is due to a tight adjustment between the growth rate and the cellular mitochondrial amount. We showed that this cellular mitochondrial amount is strictly controlled by mitochondrial biogenesis.

Mitochondrial oxidative phosphorylation is regulated by fructose 1,6-bisphosphate. A possible role in Crabtree effect induction?

In numerous cell types, tumoral cells, proliferating cells, bacteria, and yeast, respiration is inhibited when high concentrations of glucose are added to the culture medium. This phenomenon has been named the "Crabtree effect." We used yeast to investigate (i) the short term event(s) associated with the Crabtree effect and (ii) a putative role of hexose phosphates in the inhibition of respiration.

Profound effects of the general anesthetic etomidate on oxidative phosphorylation without effects on their yield.

We investigated the effects of the general anesthetic Etomidate on oxidative phosphorylation in isolated rat liver mitochondria. The study of each electron transfer site shows that there is an inhibition: mainly at complex I but also, to a lesser extent, at complex III. Moreover, with succinate as substrate, the increase in non-phosphorylating respiration is accompanied by a decrease in DeltaPsi.

Growth yield homeostasis in respiring yeast is due to a strict mitochondrial content adjustment.

In living cells, growth is the result of coupling between substrate catabolism and multiple metabolic processes taking place during net biomass formation and cell property maintenance. A crucial parameter for growth description is its yield, i.e.

The yeast cAMP protein kinase Tpk3p is involved in the regulation of mitochondrial enzymatic content during growth.

During aerobic cell growth, mitochondria must meet energy demand either by adjusting cellular mitochondrial content or by adjusting ATP production flux, allowing a constant growth yield. On respiratory substrate, the Ras/cAMP pathway has been shown to be involved in this process in the yeast Saccharomyces cerevisiae. We show that of the three cAMP protein kinase catalytic subunits, Tpk3p is the one specifically involved in the regulation of cellular mitochondrial content when energy demand decreases.

Competition of electrons to enter the respiratory chain: a new regulatory mechanism of oxidative metabolism in Saccharomyces cerevisiae.

In the yeast Saccharomyces cerevisiae, the most important systems for conveying excess cytosolic NADH to the mitochondrial respiratory chain are the external NADH dehydrogenases (Nde1p and Nde2p) and the glycerol-3-phosphate dehydrogenase shuttle. In the latter system, NADH is oxidized to NAD+ and dihydroxyacetone phosphate is reduced to glycerol 3-phosphate by the cytosolic Gpd1p. Subsequently, glycerol 3-phosphate donates electrons to the respiratory chain via mitochondrial glycerol-3-phosphate dehydrogenase (Gut2p).

Organization and regulation of the cytosolic NADH metabolism in the yeast Saccharomyces cerevisiae.

Keeping a cytosolic redox balance is a prerequisite for living cells in order to maintain a metabolic activity and enable growth. During growth of Saccharomyces cerevisiae, an excess of NADH is generated in the cytosol. Aerobically, it has been shown that the external NADH dehydrogenase, Nde1p and Nde2p, as well as the glycerol-3-phosphate dehydrogenase shuttle, comprising the cytoplasmic glycerol-3-phosphate dehydrogenase, Gpdlp, and the mitochondrial glycerol-3-phosphate dehydrogenase, Gut2p, are the most important mechanisms for mitochondrial oxidation of cytosolic NADH.

NADH is specifically channeled through the mitochondrial porin channel in Saccharomyces cerevisiae.

In many kinds of permeabilized cells, the restriction of metabolite diffusion by a mitochondrial porin "closed state" has been shown to control the respiration rate. However, since in isolated mitochondria the porin appears to be always "open," the physiological relevance of a putative regulation via this channel status is now a subject of discussion. In Saccharomyces cerevisiae, in which some of the NADH dehydrogenase active sites are facing the intermembrane space, this regulatory mechanism might play an important role for the regulation of the cytosolic redox status.

Kinetic regulation of the mitochondrial glycerol-3-phosphate dehydrogenase by the external NADH dehydrogenase in Saccharomyces cerevisiae.

In the yeast Saccharomyces cerevisiae, the two most important systems for conveying excess cytosolic NADH to the mitochondrial respiratory chain are external NADH dehydrogenase (Nde1p/Nde2p) and the glycerol-3-phosphate dehydrogenase shuttle. In the latter system, NADH is oxidized to NAD+ and dihydroxyacetone phosphate is reduced to glycerol 3-phosphate by the cytosolic Gpd1p; glycerol 3-phosphate gives two electrons to the respiratory chain via mitochondrial glycerol-3-phosphate dehydrogenase (Gut2p)-regenerating dihydroxyacetone phosphate. Both Nde1p/Nde2p and Gut2p are located in the inner mitochondrial membrane with catalytic sites facing the intermembranal space.

Thyroid status is a key regulator of both flux and efficiency of oxidative phosphorylation in rat hepatocytes.

Thyroid status is crucial in energy homeostasis, but despite extensive studies the actual mechanism by which it regulates mitochondrial respiration and ATP synthesis is still unclear. We studied oxidative phosphorylation in both intact liver cells and isolated mitochondria from in vivo models of severe not life threatening hyper- and hypothyroidism. Thyroid status correlated with cellular and mitochondrial oxygen consumption rates as well as with maximal mitochondrial ATP production.

Dimethylbiguanide inhibits cell respiration via an indirect effect targeted on the respiratory chain complex I.

We report here a new mitochondrial regulation occurring only in intact cells. We have investigated the effects of dimethylbiguanide on isolated rat hepatocytes, permeabilized hepatocytes, and isolated liver mitochondria. Addition of dimethylbiguanide decreased oxygen consumption and mitochondrial membrane potential only in intact cells but not in permeabilized hepatocytes or isolated mitochondria.

Yeast mitochondrial metabolism: from in vitro to in situ quantitative study.

In this work, we first compared yeast mitochondrial oxidative metabolism at different levels of organization: whole cells (C), spheroplasts (S), permeabilized spheroplasts (PS) or isolated mitochondria (M). At present, S are more suitable for use than C for biochemical techniques such as fast extraction of metabolites and permeabilization. We show here that respiratory rates of S with various substrates are similar to C, which demonstrate that they are adapted to yeast bioenergetic studies.

Mechanisms of inhibition and uncoupling of respiration in isolated rat liver mitochondria by the general anesthetic 2,6-diisopropylphenol.

We investigated the effects of 2,6-diisopropylphenol on oxidative phosphorylation of isolated rat liver mitochondria. Diisopropylphenol strongly inhibits state-3 and uncoupled respiratory rates, when glutamate and malate are the substrates, as a direct consequence of the limitation of electron transfer at the level of complex I. In addition, diisopropylphenol acts as an uncoupler in non-phosphorylating mitochondria, which leads to an increase in respiratory rate and a large decrease in proton-motive force.

Temperature dependence of the coupling efficiency of rat liver oxidative phosphorylation: role of adenine nucleotide translocator.

We have reinvestigated the temperature dependence of the coupling efficiency of energy conversion in isolated rat liver mitochondria. We observed that respiratory control increased with temperature. Moreover, in the same conditions, the ATP/O ratio increased.

Relationships between age-dependent changes in the effect of almitrine on H(+)-ATPase/ATPsynthase and the pattern of membrane fatty acid composition.

The effects of almitrine on ATPase/ATPsynthase previously described in beef heart mitochondria (Rigoulet et al. (1990) Biochim. Biophys.

Thiobarbituric acid-reactive material content and enzymatic protection against peroxidative damage during the course of cryogenic rabbit brain edema.

The relationship between free radicals reactions and the cell detoxifying system was investigated during the development of brain edema following a cryogenic lesion in the rabbit cerebral cortex. The amount of TBA-reactive material present six hours after freezing was less than in the controls, then increased at 48 and 96 hours. The activity of superoxide dismutase (SOD) decreased 6 hours post-injury; at the same time, we observed a stimulation of catalase activity.

Relation between epileptic activities and edema formation after cryogenic injury in the rabbit.

Although cryogenic lesions have been widely used as experimental models of either focal epilepsy or focal BE, the relation between these two disorders has never been discussed. In these experiments EA and gray matter edema were correlatively investigated during the early phase (1 to 23 hr) after a cryogenic lesion in the rabbit. Indexes were developed to allow a quantitative assessment of EA.

Membrane damage in acute brain trauma.

Among a number of biochemical disturbances occurring in the acute phase of brain insults, the destruction of membrane phospholipids and its consequences on the function of membrane-bound proteins is likely to be one of the most important. In the cryogenic type of injury which is classically considered as a relevant animal model of brain contusive lesions in human traumatology, the initial attack of membranes could consist in a peroxidative damage triggered by blood ferrous compounds. This in turn would lead to an activation of phospholipase A2.

Redistribution of the flux-control coefficients in mitochondrial oxidative phosphorylations in the course of brain edema.

This work describes the control exerted by dicarboxylate carrier and succinate dehydrogenase activities on the oxidative phosphorylations in rabbit brain mitochondria as an edema develops. Vasogenic edema leads to an uncompetitive inhibition of succinate dehydrogenase activity and to a large decrease of oxidative phosphorylations linked to succinate utilisation. Naftidrofuryl treatment in vivo restores both a high succinate dehydrogenase activity and a normal respiratory rate.

Relationship between epileptic activity and edema formation in the acute phase of cryogenic lesion.

Following cryogenic lesions of the brain in the rabbit, ictal activity appears within min with a maximum at 2 h. Brain edema increases rapidly between 2-4 h with a maximum at 8 h. The glutamate concentration reaches 209% of control in the perilesional area at 2 h and the time course of glutamate/GABA ratio parallels the time course of epileptic activity.

Effects of CDP-choline on phospholipase A2 and cholinephosphotransferase activities following a cryogenic brain injury in the rabbit.

Within the tissue surrounding the necrotic lesion, following a cryogenic injury of the brain, there is a definite activation of phospholipase A2 (at 2 and 4 hr post lesion) that accounts, at least in part, for the phospholipid breakdown. There is also an activation of cholinephosphotransferase (at 2 hr post lesion) that may correspond to an early process of phospholipid resynthesis. Oral CDP-choline in this model is able to completely inhibit the activation of phospholipase A2, but has no detectable effect on cholinephosphotransferase activity.

Na+-K+-ATPase activity of glial, neuronal, and synaptosomal enriched fractions from normal and freezing-injured rabbit cerebral cortex.

This paper investigates the kinetic parameters of Na+-K+-ATPase in glial, neuronal, and synaptosomal enriched fractions isolated from rabbit cerebral cortex. Under normal conditions, kinetic parameters-Vmax and KK+0.5- of Na+-K+-ATPase are the same in the three fractions, suggesting that this enzyme behaves as the same molecular entity.