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101.
The EPR signals of oxidized and partially reduced cytochrome oxidase have been studied at pH 6.4, 7.4, and 8.4. Isolated cytochrome oxidase in both non-ionic detergent solution and in phospholipid vesicles has been used in reductive titrations with ferrocytochrome c.The g values of the low- and high-field parts of the low-spin heme signal in oxidized cytochrome oxidase are shown to be pH dependent. In reductive titrations, low-spin heme signals at g 2.6 as well as rhombic and nearly axial high-spin heme signals are found at pH 8.4, while the only heme signals appearing at pH 6.4 are two nearly axial g 6 signals. This pH dependence is shifted in the vesicles.The g 2.6 signals formed in titrations with ferrocytochrome c at pH 8.4 correspond maximally to 0.25–0.35 heme per functional unit (aa3) of cytochrome oxidase in detergent solution and to 0.22 heme in vesicle oxidase. The total amount of high-spin heme signals at g 6 found in partially reduced enzyme is 0.45–0.6 at pH 6.4 and 0.1–0.2 at pH 8.4. In titrations of cytochrome oxidase in detergent solution the g 1.45 and g 2 signals disappear with fewer equivalents of ferrocytochrome c added at pH 8.4 compared to pH 6.4.The results indicate that the environment of the hemes varies with the pH. One change is interpreted as cytochrome a3 being converted from a high-spin to a low-spin form when the pH is increased. Possibly this transition is related to a change of a liganded H2O to OH? with a concomitant decrease of the redox potential. Oxidase in phosphatidylcholine vesicles is found to behave as if it experiences a pH, one unit lower than that of the medium. 相似文献
102.
103.
N. Roth-Bejerano 《Planta》1980,149(3):252-256
The attachment of glycolate oxidase to the peroxisomal fraction derived from etiolated barley leaves (Hordeum vulgare L. cr. Dvir) is affected by light. The effect of red irradiation is reversed by subsequent far-red irradiation, indicating the involvement of phytochrome. This phytochrome effect is assumed to be related to phytochrome binding. Indeed, prevention by filipin (1.2·10-6 mol g-1 f wt) or cholesterol of phytochrome binding to membranes abolishes the effect of light on the interaction between glycolate oxidase and the peroxisomal fraction. Glycolate oxidase binding is affected by addition of quasi-ionophores such as gramicidin and filipin at a concentration of 0.6·10-3 mol g-1 f wt. This fact indicates that peroxisome-glycolate oxidase interaction may be affected by membrane potential. Since both ion transport and membrane potential are known to be affected by phytochrome, it is proposed that phytochrome acts in the light-induced modulation of glycolate oxidase attachment as a quasi-ionophore.Abbreviations GO
glycolate oxidase
- Pr and Pfr
phytochrome forms absorbing in red and far-red, respectively
- R and F
red and far-red irradiation
- Cumulative 20 Kp
20,000 g pellet obtained by centrifugation of the crude extract
- 1 Kp
1,000 g pellet
- 20 Kp
20,000 g pellet, obtained by centrifugation of 1 Kp supernatant
- 1 Kp, 20 Kp and cumulative 20 Kp
pellets obtained after density centrifugation through a sucrose cushion 相似文献
104.
The reduction of cytochrome c by beef liver sulfite oxidase was found to be strongly inhibited by high ionic strength, indicating the importance of electrostatic interactions to the reaction. The reaction rates of sulfite oxidase with singly trifluoroacetylated or trifluoromethylphenylcarbamylated cytochrome c derivatives were studied to determine the role of individual lysines in the reaction. The reaction rate was decreased by modification of the lysines immediately surrounding the heme crevice, the decreases following the order: Lys 13 > Lys 25 Lys 79 ≈ Lys 87 > Lys 8 ≈ Lys 27 ≈ Lys 72. Modification of lysines 22, 55, 88, 99, and 100 had no effect on the reaction rate. These results indicate that the interaction site on cytochrome c for sulfite oxidase is at the heme crevice region, and overlaps considerable with that for cytochrome oxidase. 相似文献
105.
106.
Robert P. Casey Clemens Broger Marcus Thelen Angelo Azzi 《Journal of bioenergetics and biomembranes》1981,13(5-6):219-228
We report here studies which characterize further the interaction ofN,N-dicyclohexylcarbodiimide with cytochromec oxidase leading to inhibition of H+ translocation by the enzyme. Further evidence is presented to show that the inhibition results from a real interaction of DCCD with the enzyme and cannot be accounted for by uncoupling and, contrary to recent criticisms, this interaction occurs specifically with subunit III of the enzyme even at relatively high inhibitor-to-enzyme stoichiometries. Use of a spin-label analogue of DCCD has enabled us to demonstrate that the carbodiimide-binding site is highly apolar and may not lie on the pathway of electron transfer.Abbreviations DCCD
N,N-dicyclohexylcarbodiimide
- NCCD
N-(2, 2, 6, 6-tetramethylpiperidyl-1-oxyl)-N-(cyclohexyl)carbodiimide
- Hepes
2-(N-2-hydroxyethylpiperazin-N-yl) ethane sulfonate
- TMPD
N,N,N,N-tetramethylphenylenediamine 相似文献
107.
P. Pasquali L. Landi L. Cabrini G. Lenaz 《Journal of bioenergetics and biomembranes》1981,13(3-4):141-148
Extraction of endogenous ubiquinone with different methods does not influence ubiquinol oxidase activity in lyophilized mitochondria in terms ofK
M, although a decrease ofV
max is sometimes observed. Experiments with submitochondrial particles from a UQ-deficient mutant ofS. cerevisiae confirm the results with UQ-depleted mitochondria and support the idea that endogenous ubiquinone is not required for the oxidation of exogenous ubiquinols by complex III. 相似文献
108.
A facultative methylotrophic bacterium was isolated from enrichment cultures containing methylamine as the sole carbon source.
It was tentatively identified as an Arthrobacter species. Extracts of cells grown on methylamine or ethylamine contained high levels of amine oxidase (E.C. 1.4.3.) activity.
Glucose- or choline-grown cells lacked this enzyme. Oxidation of primary amines by the enzyme resulted in the formation of
H2O2; as a consequence high levels of catalase were present in methylamine-and ethylamine-grown cells. The significance of catalase
in vivo was demonstrated by addition of 20 mM aminotriazole (a catalase inhibitor) to exponentially growing cells. This completely
blocked growth on methylamine whereas growth on glucose was hardly affected. Cytochemical studies showed that methylamine-dependent
H2O2 production mainly occurred on invaginations of the cytoplasmic membrane. Assimilation of formaldehyde which is generated
during methylamine oxidation was by the FBP variant of the RuMP cycle of formaldehyde fixation. The absence of NAD-dependent
formaldehyde and formate dehydrogenases indicated the operation of a non-linear oxidation sequence for formal-dehyde via hexulose
phosphate synthase. Enzyme profiles of the organism grown on various substrates suggested that the synthesis of amine oxidase,
catalase and the enzymes of the RuMP cycle is not under coordinate control. 相似文献
109.
Catechol oxidase was extracted from an acetone powder prepared from green olive. The enzyme was purified 240-fold by ammonium sulphate fractionation followed by ion exchange chromatography and gel filtration. The enzyme was characterized by substrate specificity and response to inhibitors. Between 7 and 9 bands having catechol oxidase activity could be detected by gel electrophoresis and electrofocusing. The purified enzyme had an estimated MW of 42 000. The enzyme was strongly inhibited by diethyldithiocarbamate. Inhibition by chloride was strongly dependent on pH. The enzyme did not oxidise monophenols. 相似文献
110.
The activity of cotyledon and embryo diamine oxidase was reduced by feeding ethrel and chloroethanol to the seedlings. The inhibitory effect of 2,4-D on the activity of enzyme in the cotyledon which may be mediated through ethylene was reversed by exposure of seeds to red light. 相似文献