| Titre : |
Studies on the sarcoplasmic reticulum (Ca²+ + Mg²+) ATPase |
| Type de document : |
texte imprimé |
| Auteurs : |
Bousselsela, Haoues, Auteur ; Thomas, E. W., Directeur de thèse |
| Editeur : |
University of Salford |
| Année de publication : |
1992 |
| Importance : |
216 f. |
| Présentation : |
ill. |
| Format : |
27 cm. |
| Note générale : |
Thèse d’État : Biologie : Salford, University of Salford : 1992
Bibliogr. f. 217 - 231 |
| Langues : |
Anglais (eng) |
| Mots-clés : |
ATPase enzyme Inhibition labelling Carbodiimides Phenolic compounds Cyclopiazonic |
| Index. décimale : |
D000992 |
| Résumé : |
Sarcoplasmic reticulum vesicles were isolated from rabbit hindleg and back muscle, yielding 120-130 mg SR protein using 200-300 g muscle, and resulting in a(Ca²+ + Mg²+)- ATPase with specific activity of 13-15 µmol/mg/min.
NCD-4 and NCD-5 were shown to be efficient as fluorescent probes of (Ca²+ + Mg²+)-ATPase.
Both carbodiimides inhibited the enzyme activity only in the absence of Ca²+ (specificially labelled vesicles): inclusion of Ca²+(1 mM) protects against inhibition.
A series of phenolic compounds showed an inhibitory action on (Ca²+ + Mg²+)-ATPase activity in the following order:
bis (2-hydroxy-3, 5-methylphenyl) methane> 3(4'-hydroxyphenyl) decane and (4'-hydroxyphenyl-n-heptylketone> 1,1 bis(4-hydroxyphenyl) cyclohexane> 4,4'-isopropylidene diphenol> bis (2-hydroxyphenyl)methane> 4,4'-hydroxybenzophenone> 4-methoxyphenyl-n-heptylketone.
No good correlation was obtained between the ability of phenols to inhibit (Ca²+ + Mg²+)-ATPase activity and the ability to perturb a Ca²+ -induced fluorescence quench leading to the suggestion that phenolic compounds affect other steps in the catalytic cycle besides those associated with Ca²+ -binding.
Cyclopiazonic acid was found to be an excellent inhibitor whereas its analogs "A" and "B" showed no effect on (Ca²+ + Mg²+)-ATPase activity: this may possibly ascribed to the high ATP concentration used in the essay.
All flavanoids examined exhibited an inhibitory effect on the (Ca²+ + Mg²+)-ATPase activity in the following order: robinetin> quercetin> luteolin> fisetin>galangin> rhamentin.
This group of compounds required for their potency a 7-hydroxyl group but in the presence of additional hydroxyl groups at 3'4' and 5' positions.
Hydroxyl groups 3 and 5 seem do not participate in the inhibitory potency of the flavonoids.
Quercetin was found to inhibit (Ca²+ + Mg²+)-ATPase activity reversibly with half-maximal inhibition at 10 µM.
This inhibition showed a pH dependence with the doses of quercetin being significantly lower at pH 8 than at pHs 7 and 6.4.
Also this inhibition can be removed by affinity chromatography on a Reactive Red-120 agarose matrix.
Irradiation with light of wavelengths> 350 nm of a mixture of (Ca²+ + Mg²+)-ATPase and quercetin led to covalent inhibition of enzyme, whose extent is related to quercetin and enzyme concentrations.
Pre-irradiation of quercetin followed by incubation at room temperature for 30 min with (Ca²+ + Mg²+)-ATPase did not inhibit enzyme.
The possibility of photolabelling of the enzyme is dicussed. |
Studies on the sarcoplasmic reticulum (Ca²+ + Mg²+) ATPase [texte imprimé] / Bousselsela, Haoues, Auteur ; Thomas, E. W., Directeur de thèse . - University of Salford, 1992 . - 216 f. : ill. ; 27 cm. Thèse d’État : Biologie : Salford, University of Salford : 1992
Bibliogr. f. 217 - 231 Langues : Anglais ( eng)
| Mots-clés : |
ATPase enzyme Inhibition labelling Carbodiimides Phenolic compounds Cyclopiazonic |
| Index. décimale : |
D000992 |
| Résumé : |
Sarcoplasmic reticulum vesicles were isolated from rabbit hindleg and back muscle, yielding 120-130 mg SR protein using 200-300 g muscle, and resulting in a(Ca²+ + Mg²+)- ATPase with specific activity of 13-15 µmol/mg/min.
NCD-4 and NCD-5 were shown to be efficient as fluorescent probes of (Ca²+ + Mg²+)-ATPase.
Both carbodiimides inhibited the enzyme activity only in the absence of Ca²+ (specificially labelled vesicles): inclusion of Ca²+(1 mM) protects against inhibition.
A series of phenolic compounds showed an inhibitory action on (Ca²+ + Mg²+)-ATPase activity in the following order:
bis (2-hydroxy-3, 5-methylphenyl) methane> 3(4'-hydroxyphenyl) decane and (4'-hydroxyphenyl-n-heptylketone> 1,1 bis(4-hydroxyphenyl) cyclohexane> 4,4'-isopropylidene diphenol> bis (2-hydroxyphenyl)methane> 4,4'-hydroxybenzophenone> 4-methoxyphenyl-n-heptylketone.
No good correlation was obtained between the ability of phenols to inhibit (Ca²+ + Mg²+)-ATPase activity and the ability to perturb a Ca²+ -induced fluorescence quench leading to the suggestion that phenolic compounds affect other steps in the catalytic cycle besides those associated with Ca²+ -binding.
Cyclopiazonic acid was found to be an excellent inhibitor whereas its analogs "A" and "B" showed no effect on (Ca²+ + Mg²+)-ATPase activity: this may possibly ascribed to the high ATP concentration used in the essay.
All flavanoids examined exhibited an inhibitory effect on the (Ca²+ + Mg²+)-ATPase activity in the following order: robinetin> quercetin> luteolin> fisetin>galangin> rhamentin.
This group of compounds required for their potency a 7-hydroxyl group but in the presence of additional hydroxyl groups at 3'4' and 5' positions.
Hydroxyl groups 3 and 5 seem do not participate in the inhibitory potency of the flavonoids.
Quercetin was found to inhibit (Ca²+ + Mg²+)-ATPase activity reversibly with half-maximal inhibition at 10 µM.
This inhibition showed a pH dependence with the doses of quercetin being significantly lower at pH 8 than at pHs 7 and 6.4.
Also this inhibition can be removed by affinity chromatography on a Reactive Red-120 agarose matrix.
Irradiation with light of wavelengths> 350 nm of a mixture of (Ca²+ + Mg²+)-ATPase and quercetin led to covalent inhibition of enzyme, whose extent is related to quercetin and enzyme concentrations.
Pre-irradiation of quercetin followed by incubation at room temperature for 30 min with (Ca²+ + Mg²+)-ATPase did not inhibit enzyme.
The possibility of photolabelling of the enzyme is dicussed. |
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