scholarly journals Stimulation of the alternative oxidase of Neurospora crassa by Nucleoside phosphates

1980 ◽  
Vol 188 (1) ◽  
pp. 141-144 ◽  
Author(s):  
J Vanderleyden ◽  
C Peeters ◽  
H Verachtert ◽  
H Bertrand
Keyword(s):  
Neurospora Crassa ◽  
Oxidase Activity ◽  
Alternative Oxidase ◽  
Purine Nucleoside ◽  
Submitochondrial Particles ◽  
Fold Stimulation ◽  
Succinate Oxidase ◽  
Stimulation Of

The alternative-oxidase-mediated succinate oxidase activity of Neurospora crassa decreases drastically when mitochondria are fractionated into submitochondrial particles or treated with deoxycholate. The activity, however, can be completely restored in the presence of nucleoside 5′-monophosphates. The purine nucleoside 5′-monophosphates are more effective than the pyrimidine homologues. 5′-GMP gives a 10-fold stimulation of the alternative-oxidase-mediated succinate oxidase activity in submitochondrial particles. A comparison is made with the results obtained earlier with Moniliella tomentosa [Hanssens & Verachtert (1976) J. Bacteriol. 125, 825–835; Vanderleyden, Van Den Eynde & Verachtert (1980) Biochem. J. 186, 309–316].

scholarly journals Substrate kinetics of the alternative oxidase of Neurospora crassa

1980 ◽  
Vol 186 (3) ◽  
pp. 1009-1011 ◽  
Author(s):  
J Vanderleyden ◽  
C Peeters ◽  
H Verachtert ◽  
H Bertrand
Keyword(s):  
Neurospora Crassa ◽  
Oxidase Activity ◽  
Alternative Oxidase ◽  
Submitochondrial Particles ◽  
Succinate Oxidation ◽  
Complex Ii ◽  
Ubiquinone Oxidoreductase ◽  
Kinetics Of ◽  
Substrate Kinetics

The kinetics of the succinate oxidation by cyanide-sensitive and cyanide-insensitive submitochondrial particles of Neurospora crassa cells suggest that both respiratory pathways use the same complex II. This is confirmed by comparing the kinetics of the reductase activities of the isolated succinate-ubiquinone oxidoreductase (complex II) of cyanide-sensitive and cyanide-insensitive cells respectively. No alternative-oxidase activity was found to be associated with the isolated complex II of cyanide-insensitive cells.

scholarly journals Immunological identification of the alternative oxidase of Neurospora crassa mitochondria.

1989 ◽  
Vol 9 (3) ◽  
pp. 1362-1364 ◽  
Author(s):  
A M Lambowitz ◽  
J R Sabourin ◽  
H Bertrand ◽  
R Nickels ◽  
L McIntosh
Keyword(s):  
Neurospora Crassa ◽  
Oxidase Activity ◽  
Hydroxamic Acid ◽  
Alternative Oxidase ◽  
Fungal Species ◽  
The Other ◽  
Wild Type ◽  
Higher Plant ◽  
Sauromatum Guttatum ◽  
Immunological Identification

Neurospora crassa mitochondria use a branched electron transport system in which one branch is a conventional cytochrome system and the other is an alternative cyanide-resistant, hydroxamic acid-sensitive oxidase that is induced when the cytochrome system is impaired. We used a monoclonal antibody to the alternative oxidase of the higher plant Sauromatum guttatum to identify a similar set of related polypeptides (Mr, 36,500 and 37,000) that was associated with the alternative oxidase activity of N. crassa mitochondria. These polypeptides were not present constitutively in the mitochondria of a wild-type N. crassa strain, but were produced in high amounts under conditions that induced alternative oxidase activity. Under the same conditions, mutants in the aod-1 gene, with one exception, produced apparently inactive alternative oxidase polypeptides, whereas mutants in the aod-2 gene failed to produce these polypeptides. The latter findings support the hypothesis that aod-1 is a structural gene for the alternative oxidase and that the aod-2 gene encodes a component that is required for induction of alternative oxidase activity. Finally, our results indicate that the alternative oxidase is highly conserved, even between plant and fungal species.

scholarly journals Phospholipid association with the bovine cardiac mitochondrial adenosine triphosphatase

1985 ◽  
Vol 225 (3) ◽  
pp. 597-608 ◽  
Author(s):  
R E Brown ◽  
R I Montgomery ◽  
P I Spach ◽  
C C Cunningham
Keyword(s):  
Phosphatidic Acid ◽  
Adenosine Triphosphatase ◽  
Specific Activity ◽  
Sodium Cholate ◽  
Mitochondrial Atpase ◽  
Cardiac Mitochondria ◽  
Submitochondrial Particles ◽  
Relative Enrichment ◽  
Fold Stimulation ◽  
Stimulation Of

The association of different phospholipids with a lipid-depleted oligomycin-sensitive ATPase from bovine cardiac mitochondria [Serrano, Kanner & Racker (1976) J. Biol. Chem. 251, 2453-2461] has been examined using three approaches. First, reconstitution of the ATPase with different synthetic diacyl phospholipids resulted in a 2-10-fold stimulation of ATPase specific activity depending upon the particular phospholipid employed. The phospholipid headgroup region displayed the following order of ATPase reactivation potential: dioleoylphosphatidylglycerol greater than dioleoylphosphatidic acid greater than dioleoylphosphatidylcholine. Furthermore, the ATPase showed higher levels of specific activity when reconstituted with dioleoyl phospholipid derivatives compared with dimyristoyl derivatives. Second, examination of the phospholipid remaining associated with the lipid-depleted ATPase upon purification showed that phosphatidylcholine, phosphatidylethanolamine, and diphosphatidylglycerol were present. No relative enrichment of any of these phospholipids (compared with their distribution in submitochondrial particles) was noted. Therefore, no preferential association between the ATPase and any one phospholipid could be found in the mitochondrial ATPase. Third, the sodium cholate-mediated phospholipid exchange procedure was employed for studying the phospholipid requirements of the ATPase. Replacement of about 50% of the mitochondrial phospholipid remaining with the lipid-depleted ATPase could be achieved utilizing either synthetic phosphatidic acid or phosphatidylcholine. Examination of the displaced mitochondrial phospholipid showed that phosphatidylcholine, phosphatidylethanolamine, and diphosphatidylglycerol were replaced with equal facility.

Immunological identification of the alternative oxidase of Neurospora crassa mitochondria

1989 ◽  
Vol 9 (3) ◽  
pp. 1362-1364
Author(s):  
A M Lambowitz ◽  
J R Sabourin ◽  
H Bertrand ◽  
R Nickels ◽  
L McIntosh
Keyword(s):  
Neurospora Crassa ◽  
Oxidase Activity ◽  
Hydroxamic Acid ◽  
Alternative Oxidase ◽  
Fungal Species ◽  
The Other ◽  
Wild Type ◽  
Higher Plant ◽  
Sauromatum Guttatum ◽  
Immunological Identification

Neurospora crassa mitochondria use a branched electron transport system in which one branch is a conventional cytochrome system and the other is an alternative cyanide-resistant, hydroxamic acid-sensitive oxidase that is induced when the cytochrome system is impaired. We used a monoclonal antibody to the alternative oxidase of the higher plant Sauromatum guttatum to identify a similar set of related polypeptides (Mr, 36,500 and 37,000) that was associated with the alternative oxidase activity of N. crassa mitochondria. These polypeptides were not present constitutively in the mitochondria of a wild-type N. crassa strain, but were produced in high amounts under conditions that induced alternative oxidase activity. Under the same conditions, mutants in the aod-1 gene, with one exception, produced apparently inactive alternative oxidase polypeptides, whereas mutants in the aod-2 gene failed to produce these polypeptides. The latter findings support the hypothesis that aod-1 is a structural gene for the alternative oxidase and that the aod-2 gene encodes a component that is required for induction of alternative oxidase activity. Finally, our results indicate that the alternative oxidase is highly conserved, even between plant and fungal species.

Mechanism of inhibition of succinate oxidase by products of oxidation of o-dihydroxycoumarins

1980 ◽  
Vol 45 (2) ◽  
pp. 641-652
Author(s):  
Petr Zbořil
Keyword(s):  
Inhibitory Activity ◽  
Oxidase Activity ◽  
Enzyme Preparation ◽  
Inhibitory Effects ◽  
Anaerobic Conditions ◽  
Long Period ◽  
Mechanism Of Inhibition ◽  
Intermediary Product ◽  
Succinate Oxidase ◽  
By Products

Semiquinone is an intermediary product of the oxidation of daphnetin (7,8-dihydroxycoumarin) and esculetin (6,7-dihydroxycoumarin) by diphenol oxidase; its concentration rapidly decreases. When the oxidation is effected by ferricytochrome c, the concentration of the semiquinone remains practically constant for a long period. Similarly, the ability of the products of daphnetin oxidation by diphenol oxidase to inhibit succinate oxidase activity in mitochondrial fragments rapidly decreases with time; the decrease is considerably slower in the case of cytochrome c. The inhibitory activity of the product decreases with time also during esculetin oxidation by ferricyanide. This indicates that the inhibitory effects must be ascribed predominantly to the semiquinone, the quinone is less efficient. The inhibition of succinate oxidase or succinate dehydrogenase was strongly decreased when the enzyme preparation of Keilin and Hartree was incubated with esculetin and ferricyanide in the presence of KCN or under anaerobic conditions. This demonstrates that the reaction of the inhibitor with the enzyme either involves subsequent oxidations or that the inhibitor preferentially reacts with the oxidized form of the sensitive component of the respiratory chain. The second alternative is very little probable since there is no correlation between the degree of inhibition and the binding of the inhibitor to mitochondrial fragments.

Kinetin-Mediated Stimulation of Accumulation of Buckwheat Flavonoids in the Dark

1983 ◽  
Vol 38 (9-10) ◽  
pp. 711-718 ◽  
Author(s):  
U. Margna ◽  
T. Vainjärv
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Flavonoid Biosynthesis ◽  
Accumulation Rates ◽  
Sharp Rise ◽  
Short Treatment ◽  
Exogenous Substrate ◽  
Percent Increase ◽  
High Level ◽  
Fold Stimulation ◽  
Stimulation Of

A short treatment of excised buckwheat cotyledons with a solution of kinetin lead to an up to 9-fold stimulation of anthocyanin biosynthesis, to an about 50 percent increase in the accumula­tion of rutin, and to an about 30 percent increase, on the average, in the accumulation of C-glycosylflavones in the treated material during its posttreatment incubation in the dark. When the treated cotyledons were incubated in a solution of ʟ--phenylalanine anthocyanin accumulation in the dark practically attained the same high level as it was observed in the illuminated cotyledons fed with exogenous ʟ--phenylalanine. In experiments with l4C-labelled L-phenylalanine kinetin induced a sharp rise in the labelling (resp. in the utilization of exogenous substrate for biosynthesis) of anthocyanins and rutin in the dark and a slight increase in the radioactivity of C-glycosylflavones. Similar labelling changes occurred in the illuminated cotyledons. However, both kinetin and light still more effectively promoted biosynthetic use of the endogenous sub­strate. As a result the relative portion of flavonoids formed from exogenous L-phenylalanine under these conditions showed a decrease as compared with the ratio of precursor use in the un­treated cotyledons. The results show that low accumulation rates of anthocyanins and other flavo­noids in the dark are conditioned by the limited access of substrate (ʟ--phenylalanine) molecules to the flavonoid enzymes lending further support to the idea that flavonoid biosynthesis is normally controlled at the substrate rather than at the enzymic level.

Swelling-activated transport of taurine in cultured gill cells of sea bass: physiological adaptation and pavement cell plasticity

2009 ◽  
Vol 296 (4) ◽  
pp. R1149-R1160 ◽  
Author(s):  
Martine Avella ◽  
Olivier Ducoudret ◽  
Didier F. Pisani ◽  
Philippe Poujeol
Keyword(s):  
Sea Bass ◽  
Physiological Adaptation ◽  
Dimensional Structure ◽  
Taurine Transport ◽  
Hypotonic Shock ◽  
Cells In Culture ◽  
Membrane Stretch ◽  
Gill Cells ◽  
Fold Stimulation ◽  
Stimulation Of

We have investigated volume-activated taurine transport and ultrastructural swelling response of sea bass gill cells in culture, assuming that euryhaline fish may have developed particularly efficient mechanisms of salinity adaptation. In vivo, when sea basses were progressively transferred from seawater to freshwater, we noticed a decrease in blood osmotic pressure. When gill cells in culture were subjected to 30% hypotonic shock, we observed a five-fold stimulation of [3H]taurine efflux. This transport was reduced by various anion channel inhibitors with the following efficiency: 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) > niflumic acid > DIDS = diphenylamine-2-carboxylic acid. With polarized gill cells in culture, the hypotonic shock produced a five-fold stimulation of apical taurine transport, whereas basolateral exit was 25 times higher. Experiments using ionomycin, thapsigargin, BAPTA-AM, or removal of extracellular calcium suggested that taurine transport was regulated by external calcium. The inhibitory effects of lanthanum and streptomycin support Ca2+ entry through mechanosensitive Ca2+ channels. Branchial cells also showed hypotonically activated anionic currents sensitive to DIDS and NPPB. Similar pharmacology and time course suggested the potential existence of a common pathway for osmosensitive taurine and Cl− efflux through volume-sensitive organic osmolyte and anion channels. A three-dimensional structure study revealed that respiratory gill cells began to swell only 15 s after hypoosmotic shock. Apical microridges showed membrane outfoldings: the cell surface became smoother with a progressive disappearance of ridges. Therefore, osmotic swelling may not actually induce membrane stretch per se, inasmuch as the microridges may provide a reserve of surface area. This work demonstrates mechanisms of functional and morphological plasticity of branchial cells during osmotic stress.

scholarly journals Deficient cyclic adenosine 3',5'-monophosphate control in mutants of two genes of Neurospora crassa.

1981 ◽  
Vol 1 (1) ◽  
pp. 1-8 ◽  
Author(s):  
M L Pall ◽  
J M Trevillyan ◽  
N Hinman
Keyword(s):  
Plasma Membrane ◽  
Membrane Potential ◽  
Cyclic Amp ◽  
Adenylate Cyclase ◽  
Neurospora Crassa ◽  
Alternative Oxidase ◽  
Oscillatory Behavior ◽  
Wild Type ◽  
Glucose Transport System ◽  
Regulated Functions

Strains of Neurospora crassa mutant in either of two genes, Crisp-1 (cr1) and Frost (fr), showed no increase of cyclic adenosine 3',5'-monophosphate (cyclic AMP) levels when subjected to several treatments which produce large increases of cyclic AMP in wild-type Neurospora. Evidently, the previously reported deficiencies of adenylate cyclase in these mutants were sufficient to block the normal increases. This fact suggests that both mutants could be used to help determine which control phenomena involve cyclic AMP and to interrupt the control of established cyclic AMP-regulated functions. Earlier studies had suggested an interdependence of the cyclic AMP level and the electric potential difference across the plasma membrane of Neurospora. Present experiments, therefore, employed several strains with the cr1 mutation to test for possible roles of cyclic AMP in recovery and oscillatory behavior of the Neurospora membrane potential. The results showed all such phenomena to be normal in the adenylate cyclase-defective strains, which demonstrates that variations of cyclic AMP are not obligatorily involved in the apparent control processes. Evidence is also presented that the induction of both glucose transport system II and the alternative oxidase do not require elevated cyclic AMP levels.

scholarly journals A comparison of the effects of NN′-dicyclohexylcarbodi-imide, oligomycin A and aurovertin on energy-linked reactions in mitochondria and submitochondrial particles

1968 ◽  
Vol 108 (3) ◽  
pp. 445-456 ◽  
Author(s):  
A. M. Roberton ◽  
Caroline T. Holloway ◽  
I G Knight ◽  
R B Beechey
Keyword(s):  
Potent Inhibitor ◽  
Atp Synthesis ◽  
Mitochondrial Fraction ◽  
Optimum Concentration ◽  
Submitochondrial Particles ◽  
Nicotinamide Nucleotide Transhydrogenase ◽  
Oligomycin A ◽  
Submitochondrial Particle ◽  
Site Of Action ◽  
Stimulation Of

1. The effects of dicyclohexylcarbodi-imide, oligomycin A and aurovertin on enzyme systems related to respiratory-chain phosphorylation were compared. Dicyclohexylcarbodi-imide and oligomycin A have very similar functional effects, giving 50% inhibition of ATP-utilizing and ATP-generating systems at concentrations below 0·8nmole/mg. of submitochondrial-particle protein. Aurovertin is a more potent inhibitor of ATP synthesis, giving 50% inhibition at 0·2nmole/mg. of protein. However, aurovertin is a less potent inhibitor of ATP-utilizing systems: the ATP-driven energy-linked nicotinamide nucleotide transhydrogenase is 50% inhibited at 3·0nmoles/mg. of protein and the ATP-driven reduction of NAD+ by succinate is 50% inhibited at 0·95nmole/mg. of protein. 2. With EDTA-particles (prepared by subjecting mitochondria to ultrasonic radiation at pH9 in the presence of 2mm-EDTA) the maximum stimulation of the ATP-driven partial reactions is effected by similar concentrations of oligomycin A and dicylcohexylcarbodi-imide, but the latter is less effective. The stimulatory effects of suboptimum concentrations of dicyclohexylcarbodi-imide and oligomycin A are additive. Aurovertin does not stimulate these reactions or interfere with the stimulation by the other inhibitors. 3. Dicyclohexylcarbodi-imide and oligomycin A stimulate the aerobic energy-linked nicotinamide nucleotide transhydrogenase of EDTA-particles, but the optimum concentration is higher than that required for the ATP-driven partial reactions. Aurovertin has no effect on this reaction. 4. The site of action of dicyclohexylcarbodi-imide is in CF0, the mitochondrial fraction that confers oligomycin sensitivity on F1 mitochondrial adenosine triphosphatase.

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