The purification and properties of the respiratory-chain reduced nicotinamide–adenine dinucleotide dehydrogenase of <i>Torulopsis utilis</i>

Tottmar, S. O. C.; Ragan, C I

doi:10.1042/bj1240853

Cited by 25 publications

(13 citation statements)

References 35 publications

(22 reference statements)

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“…In this context it may be noted that the detergent-solubilized NADH dehydrogenase (i.e. NADH ferricyanide reductase) of T. utilis mitochondria co-purifies with cytochrome aa [9] .…”

Section: Discussionmentioning

confidence: 99%

Mitochondrial effects of copper‐limited growth of Torulopsis utilis

Light¹

1972

FEBS Letters

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“…In this context it may be noted that the detergent-solubilized NADH dehydrogenase (i.e. NADH ferricyanide reductase) of T. utilis mitochondria co-purifies with cytochrome aa [9] .…”

Section: Discussionmentioning

confidence: 99%

Mitochondrial effects of copper‐limited growth of Torulopsis utilis

Light¹

1972

FEBS Letters

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“…For some purified eukaryotic, non-mammalian enzymes the Fe:FMN ratio's are 27.5 (Torulopsis utilis (Tottmar and Ragan 1971)), 28-38 (N. crassa (Schulte et al 1998)), 33.2 (Pichia pastoris (Bridges et al 2009)) and 30.4 (Pichia angusta (Bridges et al 2009)). The values for the purified Y. lipolytica enzyme have not been reported (Djafarzadeh et al 2000;Kashani-Poor et al 2001a).…”

Section: Quantitative Aspectsmentioning

confidence: 99%

The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited

Albracht

2010

J Bioenerg Biomembr

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The first purification of bovine NADH:ubiquinone oxidoreductase (Complex I) was reported nearly half a century ago (Hatefi et al. J Biol Chem 237:1676-1680, 1962. The pathway of electron-transfer through the enzyme is still under debate. A major obstacle is the assignment of EPR signals to the individual iron-sulfur clusters in the subunits. The preceding paper described a working model based on the kinetics with NADPH. This model is at variance with current views in the field. The present paper provides a critical overview on the possible causes for the discrepancies. It is concluded that the stability of all purified preparations described thus far, including Hatefi's Complex I, is compromised due to removal of the enzyme from the protective membrane environment. In addition, most preparations described during the last two decades are purified by methods involving synthetic detergents and column chromatography. This results in delipidation, loss of endogenous quinones and loss of reactions with (artificial) quinones in a rotenone-sensitive way. The Fe:FMN ratio's indicate that FMN-a is absent, but that all Fe-S clusters may be present. In contrast to the situation in bovine SMP and Hatefi's Complex I, three of the six expected [4Fe-4S] clusters are not detected in EPR spectra. Qualitatively, the overall EPR lineshape of the remaining three cubane signals may seem similar to that of Hatefi's Complex I, but quantitatively it is not. It is further proposed that point mutations in any of the TYKY, PSST, 49-kDa or 30-kDa subunits, considered to make up the delicate structural heart of Complex I, may have unpredictable effects on any of the other subunits of this quartet. The fact that most point mutations led to inactive enzymes makes a correct interpretation of such mutations even more ambiguous. In none of the Complex-Icontaining membrane preparations from non-bovine origin, the pH dependencies of the NAD(P)H→O 2 reactions and the pH-dependent reduction kinetics of the Fe-S clusters with NADPH have been determined. This excludes a proper discussion on the absence or presence of FMN-a in native Complex I from other organisms.

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“…2), whereas the NADPH dehydrogenase activity was about equally distributed over the inside and outside of the mitochondrial membrane. According to Tottmar & Ragan (1971) and Mackler et al (1981) the internal NADH dehydrogenase in yeasts cannot react,with NADPH. Hence the existence of internal NADPH dehydrogenase activity in C. utilis suggests that inside the mitochondrion two distinct dehydrogenases may be present for the oxidation of NADH and NADPH.…”

Section: Discussionmentioning

confidence: 99%

Oxidation of NADH and NADPH by Mitochondria from the Yeast Candida utilis

et al. 1985

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Mitochondria were isolated from Candida utilis CBS 621 grown in carbon-limited continuous cultures on glucose, gluconate, xylose, ethanol or acetate as the carbon source and ammonia or nitrate as the nitrogen source. In all cases mitochondria were isolated which could oxidize exogenous NADH and NADPH via a cyanide-and antimycin A-sensitive but rotenoneinsensitive respiratory chain, Oxidation of NADH and NADPH was coupled to energy conservation as evidenced by high respiratory control values. Different respiratory control values of mitochondria with NADH and NADPH as well as variations in the ratio of NADH and NADPH oxidase activities indicate that separate systems exist for the oxidation of exogenous redox equivalents by mitochondria of C. utilis.Variation of the NADPH requirement for biomass formation by applying different growth conditions did not result in significant changes in NADPH oxidase activities of mitochondria. It is concluded that in C. utilis NADPH can be used in dissimilatory processes for the generation of ATP. I N T R O D U C T I O NNADPH is an essential reductant in anabolic processes. Enzyme studies have revealed that the hexose monophosphate (HMP) pathway and possibly NADP+-linked isocitrate dehydrogenase are the major sources of NADPH in the yeast Candida utilis (Bruinenberg et al., 1983 b). All available evidence indicates that the organism is unable to interconvert NADH and NADPH via transhydrogenase or analogous enzyme systems (Bruinenberg et al., 1983a, b). In theoretical calculations of the NADPH requirement for biomass formiit ion it was demonstrated that for growth with glucose as the carbon source and ammoniuiii as the nitrogen source, depending on the contribution of the NADP+-dependent isocitrate dchydrogenase, 2 to 7% of the glucose metabolized has to be oxidized in the HMP pathway to meet the NADPH requirement for biomass formation (Bruinenberg et al., 1983 a). However, radiorespirometric studies have revealed that in C. utilis approximately 30 to 50% of the glucose is metabolized via the HMP pathway (Mian et al., 1974). Also in other yeasts (Suomaliiinen & Oura, 1971) the contribution of the HMP pathway to glucose metabolism may be considerably higher than the calculated minimum.Activities of the HMP pathway exceeding the theoretical minimum point to a mechanism for dissimilatory oxidation of NADPH. Indeed it was found that mitochondria o r submitochondrial particles from several yeasts and moulds oxidize NADPH (Schuurmans Stekhoven, 1966;Djavadi et al., 1980;Schwitzguebel & Palmer, 1981). In contrast.to the situation for NADH, information on the mechanism of NADPH oxidation by yeast mitochondria is limited. It was therefore decided to study qualitative and quantitative aspects of the oxidation of NADPH by yeast mitochondria in more detail.Mitochondria] oxidation of NADPH in catabolic processes counteracts its consumption in 0001-2312 0 1985 SGM

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The purification and properties of the respiratory-chain reduced nicotinamide–adenine dinucleotide dehydrogenase of Torulopsis utilis

Cited by 25 publications

References 35 publications

Mitochondrial effects of copper‐limited growth of Torulopsis utilis

Mitochondrial effects of copper‐limited growth of Torulopsis utilis

The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited

Oxidation of NADH and NADPH by Mitochondria from the Yeast Candida utilis

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