Immunoprecipitation of cyanide-insensitive alternative oxidase activity from solubilized Arum maculatum sub-mitochondrial particles

1986 ◽  
Vol 14 (5) ◽  
pp. 895-895 ◽  
Author(s):  
M. C. WALSH ◽  
I. R. COTTINGHAM ◽  
A. L. MOORE
Keyword(s):  
Oxidase Activity ◽  
Alternative Oxidase ◽  
Arum Maculatum

scholarly journals Kinetic analysis of the mitochondrial quinol-oxidizing enzymes during development of thermogenesis in Arum maculatum L

1996 ◽  
Vol 317 (1) ◽  
pp. 313-319 ◽  
Author(s):  
Graeme R. LEACH ◽  
Klaas KRAB ◽  
David G. WHITEHOUSE ◽  
Anthony L. MOORE
Keyword(s):  
Respiratory Rate ◽  
Succinate Dehydrogenase ◽  
Oxidase Activity ◽  
Alternative Oxidase ◽  
Alternative Pathway ◽  
Respiratory Control ◽  
Stage Of Development ◽  
Activation Level ◽  
Cytochrome Pathway ◽  
Arum Maculatum

The dependence of the rate of oxygen uptake upon the ubiquinone (Q)-pool reduction level in mitochondria isolated during the development of thermogenesis of Arum maculatum spadices has been investigated. At the α-stage of development, the respiratory rate was linearly dependent upon the reduction level of the Q-pool (Qr) both under state-3 and -4 conditions. Progression through the β/γ to the Δ-stage resulted in a non-linear dependence of the state-4 rate on Qr. In the Δ-stage of development, both state-3 and -4 respiratory rates were linearly dependent upon Qr due to a shift in the engagement of the alternative oxidase to lower levels of Qr. Western blot analysis revealed that increased alternative oxidase activity could be correlated with expression of a 35 kDa protein. Respiratory control was only observed with mitochondria in the α-stage of development. At the β/γ-stage of development, the addition of ADP resulted in a significant oxidation of the Q-pool which was accompanied by a decrease in the respiratory rate. This was due either to decreased contribution of the alternative pathway to the overall respiratory rate under state 3 or by deactivation of succinate dehydrogenase activity by ADP. Cold-storage of the spadices at the β-stage of development led to increased activity of both the cytochrome pathway and succinate dehydrogenase, without any change in alternative oxidase activity. Results are discussed in terms of how changes in the activation level of the alternative oxidase and succinate dehydrogenase influence the activity and engagement of the quinol-oxidizing pathways during the development of thermogenesis in A. maculatum.

scholarly journals Solubilization of the alternative oxidase of cuckoo-pint (Arum maculatum) mitochondria. Stimulation by high concentrations of ions and effects of specific inhibitors

1985 ◽  
Vol 228 (2) ◽  
pp. 309-318 ◽  
Author(s):  
C J Kay ◽  
J M Palmer
Keyword(s):  
Fatty Acids ◽  
Oxidase Activity ◽  
Alternative Oxidase ◽  
Alternative Pathway ◽  
Reduced Form ◽  
Quinol Oxidase ◽  
Low Concentrations ◽  
Competitive Inhibitors ◽  
High Concentrations ◽  
Arum Maculatum

Selective solubilization of cyanide- and antimycin-insensitive duroquinol oxidase activity from cuckoo-pint (Arum maculatum) mitochondria was achieved using taurocholate. Inhibitor-sensitivities and water-forming DQH2 (tetramethyl-p-hydroquinone, reduced form): O2 stoichiometry were the same for the alternative oxidase of intact Arum mitochondria. Cyanide-insensitive oxidation of DQH2 by intact and solubilized mitochondria was stimulated by up to four-fold by high concentrations of anions high in the Hofmeister series, such as phosphate, sulphate or citrate. Optimal (0.7 M) sodium citrate increased Vmax. for DQH2 oxidation by the solubilized preparation from 450 to 2400 nmol of O2 X min-1 X mg of protein-1 and decreased the apparent Km for DQH2 from 0.53 to 0.38 mM. Inhibition of solubilized DQH2 oxidase activity by CLAM (m-chlorobenzhydroxamic acid) and SHAM (salicylhydroxamic acid) was mixed competitive/non-competitive, with apparent inhibition constants for CLAM of 25 microM (Ki) and 81 microM (KI) and for SHAM of 53 microM (Ki) and 490 microM (KI). Propyl gallate and UHDBT were non-competitive inhibitors with respect to DQH2 (apparent Ki = 0.3 microM and 12 nM respectively). Low concentrations of C18 fatty acids selectively inhibited cyanide-insensitive oxidation by intact and solubilized mitochondria, and inhibition was reversed by 1% (w/v) bovine serum albumin. Inhibition was competitive with DQH2, suggesting that fatty acids interfere reversably with the binding of DQH2 to the oxidase. These results tend to support the view that quinol oxidation by the alternative pathway of Arum maculatum mitochondria is catalysed by a quinol oxidase protein, rather than by a non-enzymic mechanism involving fatty acid peroxidative reaction. [Rustin, Dupont & Lance (1983) Trends Biochem. Sci. 8, 155-157; (1983) Arch. Biochem. Biophys. 225, 630-639].

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].

Characterization of an alternative oxidase activity ofHistoplasma capsulatum

Yeast ◽  
2003 ◽  
Vol 20 (5) ◽  
pp. 381-388 ◽  
Author(s):  
Clayton H. Johnson ◽  
Jonathan T. Prigge ◽  
Aaron D. Warren ◽  
Joan E. McEwen
Keyword(s):  
Oxidase Activity ◽  
Alternative Oxidase

scholarly journals Regulation of Alternative Oxidase Activity by Pyruvate in Soybean Mitochondria

1994 ◽  
Vol 106 (4) ◽  
pp. 1421-1427 ◽  
Author(s):  
D. A. Day ◽  
A. H. Millar ◽  
J. T. Wiskich ◽  
J. Whelan
Keyword(s):  
Oxidase Activity ◽  
Alternative Oxidase

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 Identification of the quinone species in cyanide-sensitive and cyanide-insensitive mitochondria of Moniliella tomentosa

1980 ◽  
Vol 192 (3) ◽  
pp. 881-885 ◽  
Author(s):  
J Vanderleyden ◽  
M Meyers ◽  
H Verachtert
Keyword(s):  
Electron Transport ◽  
Oxidase Activity ◽  
Alternative Oxidase ◽  
Transport Pathways ◽  
Naturally Occurring

Moniliella tomentosa was investigated for the presence of different quinones that might be involved in the cyanide-sensitive and/or cyanide-insensitive electron-transport pathways. The naturally occurring quinone in Moniliella tomentosa was found to be ubiquinone-45. Other quinone species could not be detected. The concentration of ubiquinone-45 in mitochondria is not related to the presence or absence of the alternative oxidase activity.

scholarly journals Degradation of mitochondrial alternative oxidase in the appendices of Arum maculatum

2020 ◽  
Vol 477 (17) ◽  
pp. 3417-3431
Author(s):  
Kikukatsu Ito ◽  
Takafumi Ogata ◽  
Takanari Seito ◽  
Yui Umekawa ◽  
Yusuke Kakizaki ◽  
...  
Keyword(s):  
Cysteine Protease ◽  
Molecular Mechanisms ◽  
Alternative Oxidase ◽  
Phoenix Dactylifera ◽  
Peptide Sequence ◽  
Cysteine Protease Inhibitor ◽  
Inner Mitochondrial Membrane ◽  
Temperature Dependent ◽  
Arum Maculatum ◽  
Specific Degradation

Cyanide-resistant alternative oxidase (AOX) is a nuclear-encoded quinol oxidase located in the inner mitochondrial membrane. Although the quality control of AOX proteins is expected to have a role in elevated respiration in mitochondria, it remains unclear whether thermogenic plants possess molecular mechanisms for the mitochondrial degradation of AOX. To better understand the mechanism of AOX turnover in mitochondria, we performed a series of in organello AOX degradation assays using mitochondria from various stages of the appendices of Arum maculatum. Our analyses clearly indicated that AOX proteins at certain stages in the appendices are degraded at 30°C, which is close to the maximum appendix temperature observed during thermogenesis. Interestingly, such temperature-dependent protease activities were specifically inhibited by E-64, a cysteine protease inhibitor. Moreover, purification and subsequent nano LC–MS/MS analyses of E-64-sensitive and DCG-04-labeled active mitochondrial protease revealed an ∼30 kDa protein with an identical partial peptide sequence to the cysteine protease 1-like protein from Phoenix dactylifera. Our data collectively suggest that AOX is a potential target for temperature-dependent E-64-sensitive cysteine protease in the appendices of A. maculatum. A possible retrograde signalling cascade mediated by specific degradation of AOX proteins and its physiological significance are discussed.

Sign in / Sign up

Export Citation Format

Share Document