scholarly journals Some properties of pyruvate and 2-oxoglutarate oxidation by blowfly flight-muscle mitochondria

1972 ◽  
Vol 127 (1) ◽  
pp. 271-283 ◽  
Author(s):  
R. G. Hansford
Keyword(s):  
Oxygen Uptake ◽  
Isocitrate Dehydrogenase ◽  
Adenine Nucleotides ◽  
Slight Reduction ◽  
Glycerol Phosphate ◽  
Pyruvate Oxidation ◽  
Low Concentrations ◽  
High Concentrations ◽  
Kinetics Of ◽  
Very High

1. High rates of state 3 pyruvate oxidation are dependent on high concentrations of inorganic phosphate and a predominance of ADP in the intramitochondrial pool of adenine nucleotides. The latter requirement is most marked at alkaline pH values, where ATP is profoundly inhibitory. 2. Addition of CaCl2 during state 4, state 3 (Chance & Williams, 1955) or uncoupled pyruvate oxidation causes a marked inhibition in the rate of oxygen uptake when low concentrations of mitochondria are employed, but may lead to an enhancement of state 4 oxygen uptake when very high concentrations of mitochondria are used. 3. These properties are consistent with the kinetics of the NAD-linked isocitrate dehydrogenase (EC 1.1.1.41) from this tissue, which is activated by isocitrate, citrate, ADP, phosphate and H+ ions, and inhibited by ATP, NADH and Ca2+. 4. Studies of the redox state of NAD and cytochrome c show that addition of ADP during pyruvate oxidation causes a slight reduction, whereas addition during glycerol phosphate oxidation causes a `classical' oxidation. Nevertheless, it is concluded that pyruvate oxidation is probably limited by the respiratory chain in state 4 and by the NAD-linked isocitrate dehydrogenase in state 3. 5. The oxidation of 2-oxoglutarate by swollen mitochondria is also stimulated by high concentrations of ADP and phosphate, and is not uncoupled by arsenate.

scholarly journals Detection of endotoxins with the Limulus test in burned and unburned mice infected with different species of gramnegative bacteria

1975 ◽  
Vol 75 (1) ◽  
pp. 99-112 ◽  
Author(s):  
R. J. Jones ◽  
E. A. Roe ◽  
R. E. Dyster
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Low Concentrations ◽  
High Concentrations ◽  
Very High ◽  
Limulus Test

SUMMARYThe Limulus test detected endotoxins in the plasma of burned and unburned mice infected with different species of gram-negative bacteria. Individual strains of different species of gram-negative bacteria produced different amounts of endotoxin in the plasma of infected mice. Plasma from mice given lethal infections showed very high concentrations of endotoxin. Low concentrations of endotoxin in the plasma were tolerated by mice but high concentrations were invariably fatal. A polyvalent pseudomonas vaccine reduced endotoxin in the plasma of mice given lethal infections of Pseudomonas aeruginosa.

scholarly journals Hexokinase Regulates Kinetics of Glucose Transport and Expression of Genes Encoding Hexose Transporters inSaccharomyces cerevisiae

2000 ◽  
Vol 182 (23) ◽  
pp. 6815-6818 ◽  
Author(s):  
Thomas Petit ◽  
Jasper A. Diderich ◽  
Arthur L. Kruckeberg ◽  
Carlos Gancedo ◽  
Karel Van Dam
Keyword(s):  
Glucose Transport ◽  
Mrna Levels ◽  
High Affinity ◽  
Expression Of Genes ◽  
Hexose Transporters ◽  
Genes Encoding ◽  
High Concentrations ◽  
Kinetics Of ◽  
Very High ◽  
Null Strain

ABSTRACT Glucose transport kinetics and mRNA levels of different glucose transporters were determined in Saccharomyces cerevisiaestrains expressing different sugar kinases. During exponential growth on glucose, a hxk2 null strain exhibited high-affinity hexose transport associated with an elevated transcription of the genesHXT2 and HXT7, encoding high-affinity transporters, and a diminished expression of the HXT1 andHXT3 genes, encoding low-affinity transporters. Deletion ofHXT7 revealed that the high-affinity component is mostly due to HXT7; however, a previously unidentified very-high-affinity component (Km = 0.19 mM) appeared to be due to other factors. Expression of genes encoding hexokinases from Schizosaccharomyces pombe orYarrowia lipolytica in a hxk1 hxk2 glk1 strain prevented derepression of the high-affinity transport system at high concentrations of glucose.

scholarly journals Cyanide-insensitive oxidation of ascorbate + NNN′N′-tetramethyl-p-phenylenediamine mixture by mung-bean (Phaseolus aureus) mitochondria. An energy-linked function

1978 ◽  
Vol 176 (1) ◽  
pp. 129-136 ◽  
Author(s):  
S B Wilson
Keyword(s):  
Oxygen Uptake ◽  
Mung Bean ◽  
Electron Flow ◽  
Plant Mitochondria ◽  
Phosphorylation Site ◽  
Antimycin A ◽  
Phaseolus Aureus ◽  
Salicylhydroxamic Acid ◽  
Low Concentrations ◽  
High Concentrations

Freshly prepared washed or purified mung-bean (Phaseolus aureus) mitochondria utilize oxygen with ascorbate/tetramethyl-p-phenylenediamine mixture as electron donor in the presence of KCN. ATP control of the oxygen uptake can be observed with very fresh mitochondria. The electron flow, which is inhibited by antimycin A, salicylhydroxamic acid or octylguanidine, takes place by reversed electron transport through phosphorylation site II and thence to oxygen through the cyanide-insensitive pathway. Oligomycin and low concentrations of uncoupler partially inhibit the oxygen uptake in a manner similar to that observed for other energy-linked functions of plant mitochondria. An antimycin A-insensitive oxygen uptake occurs if high concentrations of uncoupler are used, indicating that the pathway of electron flow has been altered. The process of cyanide-insensitive ascorbate oxidation is self-starting, and, since it occurs in the presence of oligomycin, it is concluded that the reaction can be energized by a single energy-conservation site associated with the cyanide-insensitive oxidase pathway.

Copper, nickel, and thallium uptake by the lichen Cladina rangiferina

1981 ◽  
Vol 59 (1) ◽  
pp. 91-100 ◽  
Author(s):  
M. A. S. Burton ◽  
P. LeSueur ◽  
K. J. Puckett
Keyword(s):  
Metal Uptake ◽  
Deionized Water ◽  
Metabolic Inhibitors ◽  
Temperature Dependent ◽  
Low Concentrations ◽  
Nickel Uptake ◽  
High Concentrations ◽  
Uptake Studies ◽  
Cladina Rangiferina ◽  
Very High

Metal uptake studies with Cladina rangiferina showed that the affinity for nickel was much lower than for copper or thallium. Nickel uptake was not decreased by the absence of light or oxygen or by pretreatment with metabolic inhibitors. Nickel uptake was not temperature dependent but was very dependent upon pH.Cation-exchange studies demonstrated that there was a stoichiometric exchange of Ni2+ for Sr2+, and Cu2+ for Sr2+. The exchange of Tl+ for Sr2+ was not stoichiometric, excess Tl+ was accumulated in relation to the Sr2+ released. The ratio of Sr2+:Tl+ exchange increased with increasing Tl+ availability from 1:9 (12.5 μmol Tl+ available/g of lichen) to 1:2 (500 μmol Tl+ available). Acid-treated lichen gave the expected exchange ratio of 1:2. Washing of the thalli with deionized water resulted in the continued loss of Tl+ from acid-treated and live C. rangiferina. Copper and nickel were not released in this manner.Increasing concentrations of copper and thallium produced a corresponding loss of potassium from the thallus. The potassium loss was initiated at low concentrations of copper and thallium whereas very high concentrations of manganese and nickel were required to bring about the same response.

scholarly journals Etude, dans le raisin, de l'activité β-glucosidase

OENO One ◽  
1988 ◽  
Vol 22 (2) ◽  
pp. 125
Author(s):  
C. Biron ◽  
Robert Cordonnier ◽  
Ollivier Glory ◽  
Ziya Günata ◽  
Jean-Claude Sapis
Keyword(s):  
Plant Material ◽  
Grape Berry ◽  
Cabernet Sauvignon ◽  
Mature Fruit ◽  
Glucosidase Activity ◽  
Low Concentrations ◽  
High Concentrations ◽  
Grape Leaf ◽  
Method Of Extraction ◽  
Very High

<p style="text-align: justify;">Dans ce travail, on étudie l'activité β-glucosidase du raisin. Une méthode d'extraction de l'enzyme à partir du matériel végétal a été mise au point et optimisée. L'activité enzymatique est mesurée spectrophotométriquement avec le paranitrophénylglucopyranoside comme substrat. Dans la baie de raisin, la β-glucosidase est concentrée dans les parties solides, pellicule et pulpe. Dans le jus, elle est en faible quantité. Dans la feuille de vigne, l'activité est élevée dans le limbe, faible dans le pétiole. Au cours de la maturation du raisin l'activité β-glucosidase augmente jusqu'à la maturité. Au-delà, jusqu'au stade de surmaturation, elle reste constante. Dans les raisins mars, la β-glucosidase est présente dans toutes les variétés étudiées, aussi bien dans les variétés aromatiques (Muscat d'Alexandrie, Muscat de Frontignan, Muscat de Hambourg) que dans celles non aromatiques (Carignan, Grenache, Cinsaut, Baroque, Cabernet-Sauvignon, Syrah, Merlot). Pour un millésime donné, l'activité β-glucosidase varie selon la variété. Les activités les plus élevées ont été rencontrées dans les Muscats, le Carignan, la Syrah et le Merlot. Elles sont environ 2,8 fois plus élevées que les activités les plus faibles. Pour une variété donnée, les niveaux d'activité varient fréquemment selon le millésime.</p><p style="text-align: justify;">+++</p><p style="text-align: justify;">β-glucosidase activity in grape was studied. A method of extraction of the enzyme from plant material was optimized and glucosidase activity measured spectrophotometrically using paranitrophenylglucopyranoside as substrate. High concentrations of β-glucosidase were found in grape berry solids (skin and pulp) and low concentrations in the juice. This distribution is similar to that of free terpenols in the same parts of the berry. The β-glucosidase content were very high in grape leaf blades and low in stems. Activity of the enzyme increased during maturation of the fruit. It was found in mature fruit of both aromatic (Muscat of Alexandria, Muscat of Frontignan, Muscat of Hamburg) and non-aromatic (Carignane, Grenache, Cinsaut, Baroque, Cabernet-Sauvignon, Sirah, Merlot) varieties. β-glucosidase activity varied according to variety for a given vintage. The highest activities (approximately 2.8 times higher than the lowest observed) were found in the differents Muscats and also in non-aromatic varieties such as Sirah, Merlot and Carignane. β-glucosidase activity in a given variety was frequently found to vary considerably from one vintage to another.</p>

Mitotic blocking agents for suspension cultures of maize 'Black Mexican Sweet' cell lines

Genome ◽  
1989 ◽  
Vol 32 (3) ◽  
pp. 475-478 ◽  
Author(s):  
J. Stadler ◽  
R. Phillips ◽  
M. Leonard
Keyword(s):  
Mitotic Index ◽  
Culture Medium ◽  
Colchicine Treatment ◽  
Mitotic Arrest ◽  
Suspension Cultures ◽  
Metaphase Arrest ◽  
Low Concentrations ◽  
High Concentrations ◽  
Very High ◽  
Black Mexican Sweet

Good metaphase arrest (25% mitotic index) in maize (Zea mays L.) 'Black Mexican Sweet' suspension cultures can be obtained with colchicine treatment alone, but only if very high concentrations (0.5%; 12.5 mM) are used. This colchicine concentration is 5- to 10-fold greater than that required for optimum mitotic arrest in cell cultures of other plant species. In contrast, we report that the herbicide amiprophos methyl (APM) is a much more efficient metaphase inhibitor for 'Black Mexican Sweet' suspension cultures than colchicine. Low concentrations (50 μm) of APM applied for 21–28 h produce a similar 25% mitotic index, and the growth-inhibiting effects of this treatment are undetectable 24 h after the removal of APM from the culture medium, APM, therefore, may be a useful agent for mitotic arrest in experiments which require survival of the treated cells. Another antitubulin herbicide, trifuralin, also was tested for ability to promote metaphase arrest in 'Black Mexican Sweet' cultures, but it proved to be ineffective.Key words: mitotic arrest, colchicine, phosphoric amide herbicide.

Yeast actin with a subdomain 4 mutation (A204C) exhibits increased pointed-end critical concentration

2007 ◽  
Vol 85 (3) ◽  
pp. 319-325 ◽  
Author(s):  
David J. Teal ◽  
John F. Dawson
Keyword(s):  
Actin Polymerization ◽  
Critical Concentration ◽  
Structural Studies ◽  
Wild Type ◽  
Low Concentrations ◽  
Engineering Strategy ◽  
High Concentrations ◽  
Pointed End ◽  
Very High

Characterizing mutants of actin that do not polymerize will advance our understanding of the mechanism of actin polymerization and will be invaluable for the production of short F-actin structures for structural studies. To circumvent the problem of expressing dominant lethal nonpolymerizing actin in yeast, we adopted a cysteine engineering strategy. Here we report the characterization of a mutant of yeast actin, AC-actin, possessing a single pointed-end mutation, A204C. Expression of this mutant in yeast results in actin-polymerization-deficient phenotypes. When copolymerized with wild-type actin, ATP–AC-actin is incorporated into filaments. ADP–AC-actin participates in the nucleation and elongation of wild-type filaments only at very high concentrations. At low concentrations, ADP–AC-actin appears to participate only in the nucleation of wild-type filaments, suggesting that Ala-204 is involved in modulating the critical concentration of the pointed end of actin.

Hormone-stimulated glycogenolysis in isolated goldfish hepatocytes

1976 ◽  
Vol 231 (1) ◽  
pp. 191-197 ◽  
Author(s):  
MJ Birnbaum ◽  
J Schultz ◽  
JN Fain
Keyword(s):  
Cyclic Amp ◽  
Blocking Agent ◽  
Ionophore A23187 ◽  
Bacterial Collagenase ◽  
Goldfish Carassius Auratus ◽  
Low Concentrations ◽  
Cyclic Amp Accumulation ◽  
The Common ◽  
High Concentrations ◽  
Very High

Hepatocytes isolated from the liver of the common goldfish Carassius auratus L. with crude bacterial collagenase maintained ATP levels for at least 2 h. Glycogenolysis was maximally activated by 1 X 10(-6) M epinephrine and 5.8 X 10(-9) M glucagon. In liver cells incubated in calcium-free buffer containing 1 mM ethylene glycol-bis-(beta-aminoethylether)-N,N'-tetraacetic acid, basal glycogenolysis was enhanced by the addition of 1-4 mM calcium but the elevation of cyclic AMP and glycogenolysis due to epinephrine was unaffected by calcium. The divalent cation ionophore A23187 did not alter basal or hormone-stimulated glycogenolysis. Isoproterenol was approximately as potent as epinephrine but phenylephrine was glycogenolytic only at very high concentrations. l-Propranolol competitively inhibited the increased glycogenolysis due to catecholamines but phentolamine was ineffective as a blocking agent. Isoproterenol and epinephrine stimulated glycogenolysis at lower concentrations than those required to elevate cyclic AMP accumulation. Phenylephrine was without effect on cyclic AMP. Propranolol competitively inhibited both epinephrine- and isoproterenol-stimulated cyclic AMP accumulation, but phentolamine did not block either response. Catecholamine-stimulated glycogenolysis in goldfish liver is apparently a beta-adrenergic effect. However, low concentrations of epinephrine enhance glycogenolysis without affecting total cyclic AMP.

scholarly journals Inhibition by local anaesthetics of adenine nucleotide translocation in rat liver mitochondria

1974 ◽  
Vol 140 (3) ◽  
pp. 413-422 ◽  
Author(s):  
Terry L. Spencer ◽  
Fyfe L. Bygrave
Keyword(s):  
Rat Liver ◽  
Protein Interactions ◽  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Local Anaesthetics ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Low Concentrations ◽  
High Concentrations ◽  
Lipid Protein Interactions

1. The mechanism of adenine nucleotide translocation in mitochondria isolated from rat liver was further examined by using the local anaesthetics procaine, butacaine, nupercaine and tetracaine as perturbators of lipid–protein interactions. Each of these compounds inhibited translocation of ADP and of ATP; butacaine was the most effective with 50% inhibition occurring at 30μm for 200μm-ATP and at 10μm for 200μm-ADP. The degree of inhibition by butacaine of both adenine nucleotides was dependent on the concentration of adenine nucleotide present; with low concentrations of adenine nucleotide, low concentrations of butacaine-stimulated translocation, but at high concentrations (greater than 50μm) low concentrations of butacaine inhibited translocation. Butacaine increased the affinity of the translocase for ATP to a value which approached that of ADP. 2. Higher concentrations of nupercaine and of tetracaine were required to inhibit translocation of both nucleotides; 50% inhibition of ATP translocation occurred at concentrations of 0.5mm and 0.8mm of these compounds respectively. The pattern of inhibition of ADP translocation by nupercaine and tetracaine was more complex than that of ATP; at very low concentrations (less than 250μm) inhibition ensued, followed by a return to almost original rates at 1mm. At higher concentrations inhibition of ADP translocation resulted. 3. That portion of ATP translocation stimulated by Ca2+ was preferentially inhibited by each of the local anaesthetics tested. In contrast, inhibition by the anaesthetics of ADP translocation was prevented by low concentrations of Ca2+. 4. The data provide further support for our hypothesis that lipid–protein interactions are important determinants in the activity of the adenine nucleotide translocase in mitochondria.

scholarly journals Kinetics of Alkaline Phosphatase from Pig Kidney. Influence of complexing agents on stability and activity

1976 ◽  
Vol 153 (2) ◽  
pp. 151-157 ◽  
Author(s):  
B P Ackermann ◽  
J Ahlers
Keyword(s):  
Alkaline Phosphatase ◽  
Metal Ion ◽  
Complexing Agents ◽  
Pig Kidney ◽  
Low Concentrations ◽  
Sequential Addition ◽  
Quantitative Examination ◽  
High Concentrations ◽  
Kinetics Of ◽  
Active Centres

Metal ion-complexing agents, like KCN, EDTA etc., inactivate alkaline phosphatase of pig kidney. This inactivation is reversible at low concentrations of the complexing agents and irreversible at high concentrations. The reversible inhibition is probably due to removal of Zn2+ ions from the active site, where they are necessary for catalytic action, whereas the irreversible inhibition results from the removal of Zn2+ ions necessary for preservation of the structure. The inactivation is pseudo-first order. It depends on the concentration, size and charge of the complexing agents. β-Glycerophosphate and Mg2+ ions protect the enzyme from inactivation by complexing agents. Quantitative examination of the effect of substrate leads to a model that is similar to the “sequential model” proposed by D.E. Koshland, G. Nemethy & D. Filmer (1966) (Biochemistry 5, 365-385) to explain allosteric behavior of enzymes. It describes the sequential addition of two substrate molecules at two active centres of the dimer enzyme. The binding of the substrate molecules is accompanied by changes in the conformation, which lead to stabilization of the enzyme against attack by complexing agents.

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