scholarly journals Mitochondrial adenosine triphosphatase of the fission yeast, Schizosaccharomyces pombe 972h-. Changes in activity and inhibitor-sensitivity in response to catabolite repression

1976 ◽  
Vol 160 (2) ◽  
pp. 335-342 ◽  
Author(s):  
D Lloyd ◽  
S W Edwards
Keyword(s):  
Atpase Activity ◽  
Schizosaccharomyces Pombe ◽  
Adenosine Triphosphatase ◽  
Specific Activity ◽  
Early Stage ◽  
Gel Filtration ◽  
Growth Cycle ◽  
Mitochondrial Atpase ◽  
Submitochondrial Particles ◽  
Fourfold Increase

1. The specific activity of mitochondrial ATPase (adenosine triphosphatase) in extracts of Schizosaccharomyces pombe decreased 2.5-fold as the glucose concentration in the growth medium decreased from 50mM to 15mM. 2. During the late exponential phase of growth, ATPase activity doubled. 3. Sensitivity to oligomycin and Dio-9 as measured by values for I50(mug of inhibitor/mg of protein giving 50% inhibition) at pH 6.8 increased sixfold and ninefold respectively during the initial decrease in ATPase activity, and this degree of sensitivity was maintained for the remainder of the growth cycle. 4. Increased sensitivity to NN′-dicyclohexylcarbodi-imide, triethyltin and venturicidin was also observed during the early stage of glucose de-repression. 5. Smaller increases in sensitivity to efrapeptin, aurovertin, 7-chloro-4-nitrobenzo-2-oxa-1,3-diaz-le, quercetin and spegazzinine also occurred. 6. The ATPase of glycerol-grown cells was less sensitive to inhibitors than that of glucose-repressed cells; change in values for I50 were not so marked during the growth cycle of cells growing with glycerol. 7. When submitochondrial particles from glycerol-grown cells were tested by passage through Sephadex G-50, a fourfold increase in activity was accompanied by increased inhibitor resistance. 8. Gel filtration of submitochondrial particles from glucose-de-repressed cells gave similar results, whereas loss of ATPase occurred in submitochondrial particles from glucose-repressed cells. 9. It is proposed that alterations in sensitivity to inhibitors at different stages of glucose derepression may be partly controlled by a naturally occuring inhibitor of ATPase. 10. The inhibitors tested may be classififed into two groups on the basis of alterations of sensitivity of the ATPase during physiological modification: (a) oligomycin, Dio-9, NN′-dicyclohexylcarbodi-imide, venturicidin and triethyltin, and (b) efrapeptin, aurovertin, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, quercetin and spegazzinine.

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.

scholarly journals Identity and origin of the ATPase activity associated with neuronal microtubules. II. Identification of a 50,000-dalton polypeptide with ATPase activity similar to F-1 ATPase from mitochondria.

1983 ◽  
Vol 96 (5) ◽  
pp. 1306-1315 ◽  
Author(s):  
D B Murphy ◽  
K T Wallis ◽  
R R Hiebsch
Keyword(s):  
Atpase Activity ◽  
Specific Activity ◽  
Gel Filtration ◽  
Membrane Vesicles ◽  
Cross Reactivity ◽  
Mitochondrial Atpase ◽  
Atpase Inhibitors ◽  
Native Gel ◽  
High Molecular Weight Proteins

We determined that the ATPase activity contained in preparations of neuronal microtubules is associated with a 50,000-dalton polypeptide by four different methods: (a) photoaffinity labeling of the pelletable ATPase fraction with [gamma-32P]-8-azido-ATP; (b) analysis of two-dimensional gels (native gel X SDS slab gel) of an ATPase fraction solubilized by treatment with dichloromethane; (c) ATPase purification by glycerol gradient sedimentation and gel filtration chromatography of a solvent-released ATPase fraction, (d) demonstration of the binding of affinity-purified antibody to the 50-kdalton polypeptide to ATPase activity in vitro. Beginning with preparations of microtubules we have purified the ATPase activity greater than 700-fold and estimate that the purified enzyme has a specific activity of 20 mumol Pi x mg-1 x min-1 and comprises 80-90% of the total ATPase activity associated with neuronal microtubules. With affinity-purified antibody we also demonstrate cross-reactivity to the 50-kdalton subunits of mitochondrial F-1 ATPase and show that the antibody specifically labels mitochondria in PtK-2 cells. Biochemical comparisons of the enzymes reveal similar but not identical subunit composition and sensitivity to mitochondrial ATPase inhibitors. These studies indicate that the principal ATPase activity associated with microtubules is not contained in high molecular weight proteins such as dynein or MAPs and support the hypothesis that the 50-kdalton ATPase is a membrane protein and may be derived from mitochondria or membrane vesicles with F-1-like ATPase activity.

scholarly journals Mixed anhydrides of nucleotides and mesitylenecarboxylic acid as new specific inhibitors of mitochondrial adenosien triphosphatase

1979 ◽  
Vol 178 (2) ◽  
pp. 339-343 ◽  
Author(s):  
I A Kozlov ◽  
M V Shalamberidze ◽  
I Y Novikova ◽  
N I Sokolova ◽  
Z A Shabarova
Keyword(s):  
Atpase Activity ◽  
Adenosine Triphosphatase ◽  
Covalent Binding ◽  
Mitochondrial Atpase ◽  
Mixed Anhydride ◽  
Submitochondrial Particles ◽  
Nucleoside Triphosphates ◽  
Mixed Anhydrides ◽  
Nucleotide Residue ◽  
Specific Inhibitors

Mixed anhydrides of nucleoside triphosphates and mesitylenecarboxylic acid inhibit soluble mitochondrial ATPase (adenosine triphosphatase), but do not inhibit ATPase of submitochondrial particles. Inhibition of soluble mitochondrial ATPase by the mixed anhydride of epsilon-ATP and mesitylenecarboxylic acid is followed by the covalent binding of one nucleotide residue to a molecule of the protein. It is suggested that this covalent binding occurs in the catalytic site of the mitochondrial ATPase. The mixed anhydride of ADP and mesitylenecarboxylic acid inhibits the ATPase activity of submitochondrial particles and has no effect on the activity of soluble mitochondrial ATPase. After separation of the submitochondrial particles from the mixed anhydride of ADP and mesitylenecarboxylic acid, their ATPase activity is restored to its original value (half-time of reactivation 3–4 min). Incubation of submitochondrial particles or soluble mitochondrial ATPase with the mixed anhydride of ADP and mesitylenecarboxylic acid results in AMP formation.

scholarly journals Kinetics of interaction of adenosine diphosphate and adenosine triphosphate with adenosine triphosphatase of bovine heart submitochondrial particles

1980 ◽  
Vol 188 (3) ◽  
pp. 807-815 ◽  
Author(s):  
E A Vasilyeva ◽  
A F Fitin ◽  
I B Minkov ◽  
A D Vinogradov
Keyword(s):  
Atpase Activity ◽  
Rate Constant ◽  
Adenosine Diphosphate ◽  
Mitochondrial Atpase ◽  
Dependent Manner ◽  
Submitochondrial Particles ◽  
Alternative Pathways ◽  
Atpase Reaction ◽  
Inhibitory Site ◽  
Km Value

The short preincubation of submitochondrial particles with low concentrations of ADP in the presence of Mg2+ results in a complete loss of their ATPase and inosine triphosphatase activities. Other nucleoside diphosphates (IDP and GDP) do not affect the ATPase activity. The ADP-inhibited ATPase can be activated in a time-dependent manner by treatment of submitochondrial particles with the enzyme converting ADP into ATP (phosphoenolpyruvate plus pyruvate kinase). The activaton is a first-order reaction with rate constant 0.2 min-1 at 25 degrees C. The rate constant of activation is increased in the presence of ATP up to 2 min-1, and this increase shows saturation kinetics with Km value equal to that for ATPase reaction itself (10(-4) M at 25 degrees C at pH 8.0). The experimental results obtained are consistent with the model where two alternative pathways of ADP dissociation from the inhibitory site of ATPase exist; one is spontaneous dissociation and the second is ATP-dependent dissociation through the formation of the ternary complex between ADP, the enzyme and ATP. ADP-induced inactivation and ATP-dependent activation of ATPase activity of submitochondrial particles is accompanied by the same directed change of their ability to catalyse the ATP-dependent reverse electron transport from succinate to NAD+. The possible implication of the model suggested is discussed in terms of functional role of the inhibitory high-affinity binding site for ADP in the mitochondrial ATPase.

scholarly journals The adenosine triphosphatase-inhibitor content of bovine heart submitochondrial particles. Influence of the inhibitor on adenosine triphosphate-dependent reactions

1977 ◽  
Vol 162 (2) ◽  
pp. 351-357 ◽  
Author(s):  
S J Ferguson ◽  
D A Harris ◽  
G K Radda
Keyword(s):  
Atpase Activity ◽  
Adenosine Triphosphatase ◽  
Type I ◽  
Type Ii ◽  
Inhibitor Protein ◽  
Isolation Method ◽  
Atpase Inhibitor ◽  
Submitochondrial Particles ◽  
Bovine Heart ◽  
Tightly Bound Nucleotides

1. The activity of the ATPase (adenosine triphosphatase) of phosphorylating particles prepared by sonication of bovine heart mitochondria in the presence of MgCl2 and ATP is influenced by the isolation method for the mitochondria used in the preparation of particles. Type-I particles, made from mitochondria isolated in a medium lacking succinate, have a lower ATPase activity than to Type-II particles, which are prepared from mitochondria isolated in a medium containing succinate. 2. Centrifugation under appropriate energized conditions increases the ATPase activity of Type-I particles almost to that of the Type-II particles. The ATPase activity of Type-II particles was only slightly stimulated by this procedure. These data are interpreted as indicating a higher content of the ATPase-inhibitor protein in the Type-I particles. 3. A comparison was made of the ATP-driven enhancement of 8-anilinonaphthalene-1-sulphonate fluorescence and the exchange of the endogenous tightly bound nucleotides of the ATPase in Type-I and Type-II particles. The effect of exogenous inhibitor protein on both these reactions was also studied. 4. The time-scale on which the inhibitor protein can exchange between ATPase molecules is discussed.

Isoform-independent heart rate-related variation in cardiac myofibrillar Ca(2+)-activated Mg(2+)-ATPase activity

1996 ◽  
Vol 270 (5) ◽  
pp. C1271-C1276 ◽  
Author(s):  
W. Rouslin ◽  
C. W. Broge
Keyword(s):  
Heart Rate ◽  
Beef Cattle ◽  
Atpase Activity ◽  
Specific Activity ◽  
Myosin Atpase ◽  
Curvilinear Relationship ◽  
Mitochondrial Atpase ◽  
Myofibrillar Atpase ◽  
Cardiac Myosin ◽  
Myofibrillar Atpase Activity

In the present study, we compared the activities of the cardiac myofibrillar Ca(2+)-activated Mg(2+)-ATPase and the content of cardiac muscle mitochondrial ATPase inhibitor protein (IF1) of several mammalian species covering broad ranges of body mass and heart rate, i.e., from beef cattle to mouse. The cardiac myofibrillar ATPase from each species was assayed over a range of pCa values at pH 7.4. While the cardiac myofibrillar ATPase from all species examined showed essentially identical Ca2+ concentration dependencies with the ATPase in each species activating steeply between pCa 6.5 and 5.5, the maximal ATPase specific activity reached varied considerably from species to species, and this variation was largely independent of the predominant cardiac myosin ATPase isoform present. Thus, while adult beef cattle, pig, dog, and rabbit all contain predominantly the slow cardiac myosin ATPase isoform the cardiac myofibrillar ATPase specific activities of these four species varied over approximately a fourfold range. Moreover, there was a fairly smooth curvilinear relationship between maximum Ca(2+)-activated myofibrillar ATPase activity and median conscious heart rate for the slow cardiac myosin ATPase-possessing species examined. This smooth continuum also extended to include two species possessing the fast cardiac myosin ATPase isoform, rat and mouse. This relationship between myofibrillar ATPase activity and heart rate that appears to be applicable to a broad range of species suggests that the myofibrillar ATPase is specifically modeled or fine-tuned to the kinetic (heart rate) demand of each species and, within slow and fast heart rate ranges, is essentially independent of myosin ATPase isoform per se. Only hearts containing predominantly the slow myosin ATPase isoform contained functional levels of IF1. Finally, while it has been reported that the ratio of myosin Ca(2+)-ATPase to actomyosin Mg(2+)-ATPase activity is a good index of the percent of the fast myosin ATPase in rabbit myofibrillar preparations, we found that this relationship may be applicable to only some species.

scholarly journals Mitochondrial adenosine triphosphatase of the fission yeast Schizosaccharomyces pombe 972h-. Changes in inhibitor sensitivities during the cell cycle indicate similarities and differences in binding sites

1977 ◽  
Vol 162 (3) ◽  
pp. 581-590 ◽  
Author(s):  
D Lloyd ◽  
S W Edwards
Keyword(s):  
Cell Cycle ◽  
Schizosaccharomyces Pombe ◽  
Binding Sites ◽  
Adenosine Triphosphatase ◽  
Specific Activity ◽  
Inhibitor Binding ◽  
Titration Curves ◽  
Catalytic Sites ◽  
Cycle Dependent ◽  
Similar Site

1. We used 11 different inhibitors of energy conservation as inhibitors of ATPase (adenosine triphosphatase) in extracts of Schizosaccharomyces pombe obtained from cells at different stages of the cell cycle. 2. All the inhibitors showed cell-cycle-dependent variations in their I50 values (microng of inhibitor/mg of protein giving 50% inhibition of inhibitor-sensitive ATPase at pH 8.6). 3. From the sensitivity profiles through the cell cycle it was concluded that: (a) oligomycin, venturicidin, triethyltin sulphate and dibutylchloromethyltin chloride all act at closely associated site(s); (b) NN'-dicyclohexylcarbodi-imide and leucinostatin both act at a similar site, which is, however, distinct from that at which other inhibitors of the membrane factor (Fo) act. 4. The variations in I50 values for efrapeptin closely followed changes in specific activity of ATPase, as would be expected for an inhibitor acting at catalytic sites; these fluctuations were different from those for aurovertin, Dio-9, 4-chloro-7-nitrobenzofurazan, quercetin and spegazzinine, all of which show different sensitivity profiles from one another. 5. Anomalous stepwise inhibitor-titration curves were obtained for spegazzinine, NN'-dicyclohexylcarbodiimide, dibutylchloromethyltin chloride and leucinostatin. 6. Possible explanations are proposed for the discontinuous expression of inhibitor-binding sites during the cell cycle.

scholarly journals Mitochondrial adenosine triphosphatase of the fission yeast, Schizosaccharomyces pombe 972h−. Changes in activity and oligomycin-sensitivity during the cell cycle of catabolite-repressed and -de-repressed cells

1977 ◽  
Vol 162 (1) ◽  
pp. 39-46 ◽  
Author(s):  
S W Edwards ◽  
D Lloyd
Keyword(s):  
Cell Cycle ◽  
Atpase Activity ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Adenosine Triphosphatase ◽  
Total Activity ◽  
Stages Of Growth ◽  
Synchronous Cultures ◽  
A Cell ◽  
In Cells

1. Changes in activity of ATPase (adenosine triphosphatase) during the cell cycle of Schizosaccharomyces pombe were analysed in cell-free extracts of cells harvested from different stages of growth of synchronous cultures and also after cell-cycle fractionation. 2. Oligomycin-sensitive ATPase oscillates in both glucose-repressed synchronous cultures and shows four maxima of activity approximately equally spaced through the cell cycle. The amplitude of the oscillations accounts for between 13 and 80% of the total activity at different times in the cell cycle. 3. Oligomycin sensitivity varies over a fourfold range at different stages of the cell cycle. 4. The periodicity of maximum oligomycin sensitivity is one-quarter of a cell cycle. 5. These results were confirmed for the first three-quarters of the cell cycle by cell-cycle fractionation. 6. In cells growing synchronously with glycerol, ATPase activity increases in a stepwise pattern, with two steps per cell cycle; the first of these occurs at 0.54 of the cell cycle and the second at 0.95. 7. These results are discussed in relation to previously obtained data on the development of mitochondrial activities during the cell cycle.

scholarly journals A method for determining the adenosine triphosphatase content of energy-transducing membranes. reaction of 4-chloro-7-nitrobenzofurazan with the adenosine triphosphatase of bovine heart submitochondrial particles

1976 ◽  
Vol 159 (2) ◽  
pp. 347-353 ◽  
Author(s):  
S J Ferguson ◽  
W J Lloyd ◽  
G K Radda
Keyword(s):  
Mitochondrial Membrane ◽  
Adenosine Triphosphatase ◽  
Plasma Membranes ◽  
Single Amino Acid ◽  
Mitochondrial Atpase ◽  
Inner Mitochondrial Membrane ◽  
Submitochondrial Particles ◽  
Bovine Heart ◽  
Single Amino Acid Residue ◽  
Membrane Bound

1. Modification of a single amino acid residue by introduction of the nitrobenzofurazan group inactivates mitochondrial ATPase (adenosine triphosphatase) when membrane-bound in submitochondrial particles. The similarity between the reactions of both membrane-bound and isolated ATPase with 4-chloro-7-nitrobenzofurazan indicates that the single essential tryosine residue identified in the isolated enzyme [Ferguson, Loyd, Lyons & Radda (1975) Eur. J. Biochem. 54, 117-126] Is also a feature of the membrane-bound ATPase. 2. A procedure is presented for estimating the ATPase content of the inner mitochondrial membrane. It is based on the specificity of the incorporation of the nitrobenzofurazan group, and the ready removal of this group by compounds that contain a thiol group. This method indicates that 8.5% of the membrane protein is ATPase. The procedure should be applicable to the titration of the energy-transducing ATPases of bacterial plasma membranes and of the thylakoid membranes of chloroplasts. 3. Combination of the data obtained on the ATPase content of the bovine heart inner mitochondrial membrane with a titration of the cytochrome bc1 complex with antimycin indicates that these two components of the membrane are present in approximately equal amounts.

scholarly journals The adenosine triphosphatase and calcium ion-transporting activities of the sarcoplasmic reticulum of developing muscle

1969 ◽  
Vol 114 (2) ◽  
pp. 161-170 ◽  
Author(s):  
D. L. Holland ◽  
S V Perry
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Atpase Activity ◽  
Adenosine Triphosphatase ◽  
Specific Activity ◽  
Sucrose Density Gradient ◽  
Maximum Activity ◽  
Longissimus Dorsi ◽  
Longissimus Dorsi Muscle ◽  
Dorsi Muscle ◽  
Coupling System

1. The ATPase (adenosine triphosphatase) specific activity and the total nitrogen content of the myofibrillar fraction per g. wet weight of rabbit longissimus dorsi muscle increased steadily during the late foetal stages and the first few weeks after birth. 2. The ATPase specific activity of the sarcoplasmic-reticular fraction isolated by a sucrose-density-gradient procedure rose to a sharp peak 8–10 days after birth and then declined to the adult value, which was about 25% of the maximum. 3. The peak in ATPase activity was a feature of the sarcoplasmic reticulum isolated from muscle, and the time at which it occurred in relation to birth was related to the degree of development and the activity pattern of the muscle. 4. The peak in ATPase activity of the sarcoplasmic reticulum occurred at an earlier age if newborn animals were made to exercise earlier than was normal. 5. The ‘extra’ ATPase associated with the sarcoplasmic reticulum and the ability to concentrate Ca2+ increased in a similar manner over the period of development studied. 6. It is postulated that the Ca2+-transport system of the sarcoplasmic reticulum consists of two components, namely the ATPase and the system coupling this enzyme to Ca2+ transport. During development the ATPase develops first and has almost reached maximum activity in the longissimus dorsi muscle of the rabbit after 8–10 days. Subsequently the activity of the coupling system rises rapidly, leading to an increase in the capacity and efficiency of Ca2+ transport.

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