Properties of the common inhibitor-binding domain in mitochondrial NADH-dehydrogenase (complex I).
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Bacterial Proteins↗
Biomedical subjects
U Brandt
Publications and source records attributed to U Brandt.
Anesthesiology and intensive care during epilepsy surgery in children and adolescents.
Thus far there are no known studies of the management of anesthesiology and intensive care in cases of implantation of subdural plates for the purpose of determining the epileptogenic areal and of subsequent epilepsy surgery. Such operative measures are still considered too risky, especially in the case of small children. The authors report on cases of 45 children and adolescents who underwent this kind of surgery between December 1992 and December 1997 in Gilead hospital. In order to judge the anesthesiological risk the children were divided into three age groups: A, 1-5 years (n = 12); B, 6-11 years (n = 14); C, 12-18 years (n = 19). In none of these groups were there complications which in retrospect would have shed a negative light on the operation.
Adjuvants, Anesthesia↗
A single external enzyme confers alternative NADH:ubiquinone oxidoreductase activity in Yarrowia lipolytica.
NADH:ubiquinone oxidoreductases catalyse the first step within the diverse pathways of mitochondrial NADH oxidation. In addition to the energy-conserving form commonly called complex I, fungi and plants contain much simpler alternative NADH:ubiquinone oxido-reductases that catalyze the same reaction but do not translocate protons across the inner mitochondrial membrane. Little is known about the distribution and function of these enzymes. We have identified YLNDH2 as the only gene encoding an alternative NADH:ubiquinone oxidoreductase (NDH2) in the obligate aerobic yeast Yarrowia lipolytica. Cells carrying a deletion of YLNDH2 were fully viable; full inhibition by piericidin A indicated that complex I activity was the sole NADH:ubiquinone oxidoreductase activity left in the deletion strains. Studies with intact mitochondria revealed that NDH2 in Y. lipolytica is oriented towards the external face of the mitochondrial inner membrane. This is in contrast to the situation seen in Saccharomyces cerevisiae, Neurospora crassa and in green plants, where internal alternative NADH:ubiquinone oxidoreductases have been reported. Phylogenetic analysis of known NADH:ubiquinone oxidoreductases suggests that during evolution conversion of an ancestral external alternative NADH:ubiquinone oxidoreductase to an internal enzyme may have paved the way for the loss of complex I in fermenting yeasts like S. cerevisiae.
Amino Acid Sequence↗
The chemistry and mechanics of ubihydroquinone oxidation at center P (Qo) of the cytochrome bc1 complex.
The emerging X-ray structures of the cytochrome bc1 complexes from bovine and chicken heart mitochondria support the protonmotive Q-cycle as the overall electron- and proton-pathway within the cytochrome bc1 complex. The energy conserving reaction within this reaction scheme is the unique bifurcation of electron flow into a high potential and a low potential pathway occurring at the ubihydroquinone-oxidation center (center P or Qo). This step is prerequisite for the 'recycling' of every second electron across the membrane onto the ubiquinone-reduction center, which results in vectorial proton translocation. It has been shown that during steady-state the step controlling this reaction is the first deprotonation of ubihydroquinone and not, as proposed earlier, the formation of a highly unstable semiquinone species. Ubiquinone has not yet been detected at the ubihydroquinone-oxidation center of the protein structures now available, but the pocket seems spacious enough to accommodate two ubiquinone molecules. This is in line with recent enzymological studies, which have shown that not only two ubiquinones, but also two inhibitor molecules can bind to center P. The most striking result from the structures is that the hydrophilic domain of the 'Rieske' protein can be found in two different positions which seem to allow electron transfer between the iron-sulfur cluster and either ubiquinone binding at center P or heme c1. This provides strong support for the 'catalytic switch' model proposed earlier based on detailed analysis of inhibitor binding to cytochrome bc1 complex in different redox states.
Animals↗
Identification and characterization of a novel 9.2-kDa membrane sector-associated protein of vacuolar proton-ATPase from chromaffin granules.
Vacuolar proton-translocating ATPase (holoATPase and free membrane sector) was isolated from bovine chromaffin granules by blue native polyacrylamide gel electrophoresis. A 5-fold excess of membrane sector over holoenzyme was determined in isolated chromaffin granule membranes. M9.2, a novel extremely hydrophobic 9.2-kDa protein comprising 80 amino acids, was detected in the membrane sector. It shows sequence and structural similarity to Vma21p, a yeast protein required for assembly of vacuolar ATPase. A second membrane sector-associated protein (M8-9) was identified and characterized by amino-terminal protein sequencing.
Amino Acid Sequence↗
Effects of mutation of residue I67 on redox-linked protonation processes in yeast cytochrome c oxidase.
We describe effects of a mutation, Ile-67-->Asn, in subunit I of yeast cytochrome c oxidase on redox-linked protonation processes within the protein. The mutation lowers the midpoint potential of haem a and weakens its pH dependency, but has little effect on the potential of haem a3. The residue is close to a conserved glutamate (Glu-243) in the crystal structure. We propose that protonation of Glu-243 is redox-linked to haem a, that Asn-167 perturbs its pK and that redox-linked protonation in this location is essential for the catalytic reactions of the binuclear centre. These proposals are discussed in terms of a 'glutamate trap' mechanism for proton translocation in the haem/copper oxidases.
Asparagine↗
Action of insulin on the surface morphology of hepatocytes: role of phosphatidylinositol 3-kinase in insulin-induced shape change of microvilli.
In previous studies we have shown that the insulin-responding glucose transporter isoform of 3T3-L1 adipocytes, GluT4, is almost completely located on microvilli. Furthermore, insulin caused the integration of these microvilli into the plasma membrane, suggesting that insulin-induced stimulation of glucose uptake may be due to the destruction of the cytoskeletal diffusion barrier formed by the actin filament bundle of the microvillar shaft regions [Lange et al. (1990) FEBS Lett. 261, 459-463; Lange et al. (1990) FEBS Lett. 276, 39-41]. Similar shape changes in microvilli were observed when the transport rates of adipocytes were modulated by glucose feeding or starvation. Here we demonstrate that the action of insulin on the surface morphology of hepatocytes is identical to that on 3T3L1 adipocytes; small and narrow microvilli on the surface of unstimulated hepatocytes were rapidly shortened and dilated on top of large domed surface areas. The aspect and mechanism of this effect are closely related to "membrane ruffling" induced by insulin and other growth factors. Pretreatment of hepatocytes with the PI 3-kinase inhibitor wortmannin (100 nM), which completely prevents transport stimulation by insulin in adipocytes and other cell types, also inhibited insulin-induced shape changes in microvilli on the hepatocyte surface. In contrast, vasopressin-induced microvillar shape changes in hepatocytes [Lange et al. (1997) Exp. Cell Res. 234, 486-497] were insensitive to wortmannin pretreatment. These findings indicate that PI 3-kinase products are necessary for stimulation of submembrane microfilament dynamics and that cytoskeletal reorganization is critically involved in insulin stimulation of transport processes. The mechanism of the insulin-induced cytoskeletal reorganization can be explained on the basis of the recent finding of Lu et al. [Biochemistry 35(1996) 14027-14034] that PI 3-kinase products exhibit much higher affinity for the profilin-actin complex than the primary products, PIP and PIP2. Thus, activated PI 3-kinase may direct a flux of profilin-actin complexes to the membrane locations of activated insulin receptors, where, due to the release of actin monomers after binding of profilactin to PI(3,4)P2 and PI(3,4,5)P3, massive actin polymerization is initiated. As a consequence, PI 3-kinase activation initiates a vectorial reorganization of the cellular actin system to membrane sites neighboring activated insulin receptors, giving rise to local membrane stress as visualized by extensive surface deformations and shortening of microvilli. In addition, extensive high-affinity binding of F-actin-barbed endcapping proteins enhances the cytoplasmic concentration of rapidly polymerizing filament ends. Consequently, the actin monomer concentration is lowered and the (cytoplasmic) pointed ends of the microvillar shaft bundle depolymerize and become shorter. The observations presented strengthen the previously postulated diffusion-barrier concept of glucose- and ion-uptake regulation and provide a mechanistic basis for explaining the action of insulin and other growth factors on transport processes across the plasma membrane.
Actins↗
Binding of MOA-stilbene to the mitochondrial cytochrome bc1 complex is affected by the protonation state of a redox-Bohr group of the 'Rieske' iron-sulfur protein.
MOA-stilbene is a specific inhibitor of the ubihydroquinone oxidation center (center P or o) of cytochrome bc1 complex. Binding of this inhibitor does not require the 'Rieske' iron-sulfur protein, but is affected by the redox-state of the cytochrome bc1 complex. We have analyzed the pH dependence of the apparent dissociation constant for MOA-stilbene. A 2.5 fold change in affinity between pH 6.0 and 9.5 was observed for oxidized bovine cytochrome bc1 complex. The pH profile could be simulated by assuming a single protonable group with pKA = 7.7. This pKA was not observed after partial or complete reduction of the enzyme or after removal of the iron-sulfur protein. We conclude that this protonable group was identical to the redox-Bohr group with the same pKA that has been reported to be associated with the 'Rieske' iron-sulfur cluster. Fully reduced cytochrome bc1 complex exhibited an additional binding site for MOA-stilbene. As this second binding site was abolished by the center P inhibitor stigmatellin, but not by antimycin, an inhibitor of ubiquinone reduction at center N, we conclude that it is also located at center P.
Animals↗
Role of deprotonation events in ubihydroquinone:cytochrome c oxidoreductase from bovine heart and yeast mitochondria.
The pH dependence of bovine and yeast cytochrome bc1 complex catalyzing electron transfer from ubi- and plastohydroquinone to cytochrome c have been analyzed. The pH dependence of the steady-state rate was found to be governed by two protonable groups, one of which (pK approximately 6.6) has to be deprotonated while the other (pK approximately 9.2) has to be protonated to allow catalysis. Using ubideuteroquinone instead of ubihydroquinone as a substrate resulted in 1.4- and 1.7-fold lower steady-state rates for the bovine and yeast enzymes, respectively. The activation energy at pH 8.0 was 33 kJ/mol for the bovine and 44 kJ/mol for the yeast enzyme and exhibited a linear decrease between pH 5.4 and 9.2. For ubihydroquinone the slope was very close to a value of -5.7 kJ/mol expected if the activation energy depended on a single deprotonation event. When plastohydroquinone was used instead, the slope more than doubled, indicating that a second deprotonation contributed to the activation barrier with this nonphysiological substrate. In contrast to previous kinetic models for the cytochrome bc1 complex, which propose that the activation barrier is associated with the formation of ubisemiquinone at the ubihydroquinone oxidation center, our results strongly suggest that the best approximation of the transition state is the singly deprotonated form of ubihydroquinone. This supports the recently proposed proton-gated charge transfer mechanism, which has control of catalysis by the first deprotonation of ubihydroquinone as one of its key features [Brandt, U. (1996) FEBS Lett. 387, 1-6]. All results reported here can be rationalized in a straightforward way based on other aspects of the same hypothesis.
Animals↗
Structural and functional analysis of deficient mutants in subunit I of cytochrome c oxidase from Saccharomyces cerevisiae.
Four point mutations in subunit I of cytochrome c oxidase from Saccharomyces cerevisiae that had been selected for respiratory incompetence but still contained spectrally detectable haem aa3 were analysed. The isolated mutant enzymes exhibited minor band shifts in their optical spectra and contained all eleven subunits. However, steady state activities were only a few percent compared to wild type enzyme. Using a comprehensive experimental approach, we first checked the integrity of the enzyme preparations and then identified the specific functional defect. The results are discussed using information from the recently solved structures of cytochrome c oxidase at 2.8 A. Mutation 167N is positioned between haem a and a conserved glutamate residue (E243). It caused a distortion of the EPR signal of haem a and shifted its midpoint potential by 54 mV to the negative. The high-resolution structure suggests that the primary reason for the low activity of the mutant enzyme could be that asparagine in position 67 might form a stable hydrogen bond to E243, which is part of a proposed proton channel. Cytochrome c oxidase isolated from mutant T316K did not meet our criteria for homogeneity and was therefore omitted from further analysis. Mutants G352V and V380M exhibited an impairment of electron transfer from haem a to a3 and ligand binding to the binuclear centre was affected. In mutant V380M also the midpoint potential of CuB was shifted by 65 mV to the positive. The results indicated for these two mutants changes primarily associated with the binuclear centre, possibly associated with an interference in the routes and/or sites of protonation which are required for stable formation of the catalytic intermediates. This interpretation is discussed in the light of the high resolution structure.
Electron Transport Complex IV↗
A new concept for risk assessment of the hazards of non-genotoxic chemicals--electronmicroscopic studies of the cell surface. Evidence for the action of lipophilic chemicals on the Ca2+ signaling system.
Recently, we presented evidence for the localization of components of the cellular Ca2+ signaling pathway in microvilli. On stimulation of this pathway, microvilli undergo characteristic morphological changes which can be detected by scanning electron microscopy (SEM) of the cell surface. Here we show that both receptor-mediated (vasopressin) and unspecific stimulation of the Ca2+ signaling system by the lipophilic tumor promoters thapsigargin (TG) and phorbolmyristateacetate (PMA) are accompanied by the same type of morphological changes of the cell surface. Since stimulated cell proliferation accelerates tumor development and sustained elevation of the intracellular Ca2+ concentrations is a precondition for stimulated cell proliferation, activated Ca2+ signaling is one possible mechanism of non-genomic tumor promotion. Using isolated rat hepatocytes we show that all tested lipophilic chemicals with known tumor promoter action, caused characteristic microvillar shape changes. On the other hand, lipophilic solvents that were used as differentiating agents in cell cultures such as dimethylsulfoxide (DMSO) and dimethylformamide also, failed to change the microvillar shapes. Instead DMSO stabilized the original appearance of microvilli. The used technique provides a convenient method for the evaluation of non-genomic carcinogenicity of chemicals prior to their industrial application.
Alkanes↗
Proton-translocation by membrane-bound NADH:ubiquinone-oxidoreductase (complex I) through redox-gated ligand conduction.
For the catalytic mechanism of proton-translocating NADH-dehydrogenase (complex I, EC 1.6.99.3) a number of hypothetical models have been proposed over the last three decades. These models are discussed in the light of recent substantial progress on the structure and function of this very complicated multiprotein complex. Only the high-potential iron-sulfur center N-2 and ubiquinone seem to contribute to the proton-translocating machinery of complex I: Based on the pH dependent midpoint potential of iron-sulfur cluster N-2 and the physical properties of ubiquinone intermediates a novel mechanism is proposed. The model builds on a series of defined chemical reactions taking place at three different ubiquinone-binding sites. Therefore, some aspects of this redox-gated ligand conduction mechanism are reminiscent to the proton-motive Q-cycle. However, its central feature is the abstraction of a proton from ubihydroquinone by a redox-Bohr group associated with iron-sulfur cluster N-2. Thus, in the proposed mechanism proton translocation is driven by a direct linkage between redox dependent protonation of iron-sulfur cluster N-2 and the redox chemistry of ubiquinone.
Animals↗
Evolutionary origin of cryptomonad microalgae: two novel chloroplast/cytosol-specific GAPDH genes as potential markers of ancestral endosymbiont and host cell components.
Cryptomonads are complex microalgae which share characteristics of chromophytes (chlorophyll c, extra pair of membranes surrounding the plastids) and rhodophytes (phycobiliproteins). Unlike chromophytes, however, they contain a small nucleus-like organelle, the nucleomorph, in the periplastidial space between the inner and outer plastid membrane pairs. These cellular characteristics led to the suggestion that cryptomonads may have originated via a eukaryote-eukaryote endosymbiosis between a phagotrophic host cell and a unicellular red alga, a hypothesis supported by rRNA phylogenies. Here we characterized cDNAs of the nuclear genes encoding chloroplast and cytosolic glyceraldehyde-3-phosphate dehydrogenases (GAPDH) from the two cryptomonads Pyrenomonas salina and Guillardia theta. Our results suggest that in cryptomonads the classic Calvin cycle GAPDH enzyme of cyanobacterial origin, GapAB, is absent and functionally replaced by a photosynthetic GapC enzyme of proteobacterial descent, GapC1. The derived GapC1 precursor contains a typical signal/transit peptide of complex structure and sequence signatures diagnostic for dual cosubstrate specificity with NADP and NAD. In addition to this novel GapC1 gene a cytosol-specific GapC2 gene of glycolytic function has been found in both cryptomonads showing conspicuous sequence similarities to animal GAPDH. The present findings support the hypothesis that the host cell component of cryptomonads may be derived from a phototrophic rather than a organotrophic cell which lost its primary plastid after receiving a secondary one. Hence, cellular compartments of endosymbiotic origin may have been lost or replaced several times in eukaryote cell evolution, while the corresponding endosymbiotic genes (e.g., GapC1) were retained, thereby increasing the chimeric potential of the nuclear genome.
Amino Acid Sequence↗
Pterulinic acid and pterulone, two novel inhibitors of NADH:ubiquinone oxidoreductase (complex I) produced by a Pterula species. I. Production, isolation and biological activities.
Pterulinic acid (1) and pterulone (2), two novel halogenated antibiotics, were isolated from fermentations of Pterula sp. 82168. Both compounds exhibited significant antifungal and weak or no cytotoxic activities. 1 and 2 are effective inhibitors of eucaryotic respiration. The target of the antibiotics resides within the mitochondrial NADH:ubiquinone oxidoreductase (complex I).
Acetates↗
Detection of the ATP-dependent nonmitochondrial calcium store in a cell surface-derived vesicle fraction from isolated rat hepatocytes.
In preceding studies, the IP3-sensitive Ca2+ store of the hamster insulinoma cell line, HIT, was detected in cell surface protrusions such as microvilli and related membrane structures [Lange, K., and Brandt, U. (1993) FEBS Lett. 320, 183-188; and (1993) FEBS Lett. 325, 205-209]. In this study, these experiments were extended on rat hepatocytes. We used the previously described shearing technique for isolating cell surface-derived vesicle fractions from freshly isolated and 48-h-cultured rat hepatocytes. As shown by Western blot analysis, these vesicles contained the hepatocyte-specific glucose transporter, GluT2, and actin, which are both typical microvillar components. Scanning electron microscopy revealed that a spherical vesicle population of uniform size (about 1 microm in diameter) originates from the hepatocyte microvilli. This vesicle fraction exhibited ATP-dependent and thapsigargin-sensitive Ca2+ storage activity with properties identical to those of the known microsomal systems and of HIT cell surface-derived vesicles, except that the ATP-dependent Ca2+ pool was insensitive to IP3. Like HIT surface vesicles, hepatocyte surface vesicles rapidly took up ATP via a 4,4'-diisocyanostilbene-2,2'-disulfonic acid (DIDS)-sensitive anion pathway. Inhibition of ATP influx into the vesicles by DIDS also completely inhibited ATP-dependent Ca2+ storage. Moreover, determination of efflux kinetics of Ca2+ from passively (in the absence of ATP) loaded vesicles revealed a La(3+)-sensitive but IP3-independent Ca2+ pathway which rapidly equilibrated intravesicular free Ca2+ with the external medium. Permeabilization of the vesicles with saponin (0.005%) opened an additional efflux pathway for Ca2+ which is not La(3+)-sensitive. However, saponin treatment of vesicles preloaded with Ca2+ in the presence of ATP did not affect the thapsigargin-sensitive vesicular Ca2+ store but only released a small portion (about 20%) of the vesicular Ca2+ that is not part of the thapsigargin-sensitive Ca2+ pool. Also, the size of the saponin-releasable Ca2+ pool was not affected by depletion of the thapsigargin-sensitive Ca2+ store. These findings indicate that hepatocyte surface vesicles are readily permeable for Ca2+ and ATP via cation and anion pathways. Consequently, Ca2+ storage into these vesicles does not occur by concentrative Ca2+ pumping but rather appears to be due to an internal, ATP-dependent mechanism of Ca2+ sequestration. The presented data are in accord with the previously reported colocalization of the ATP-dependent Ca2+ store and its functionally coupled, store-regulated Ca2+ influx pathway in special cell surface organelles, the microvilli.
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
Calcium storage and release properties of F-actin: evidence for the involvement of F-actin in cellular calcium signaling.
Preceding studies have shown that the bulk of the ATP-dependent, inositol 1,4,5-trisphosphate (IP3)-sensitive Ca2+ store of hamster insulinoma (HIT) cells is located in microvilli on the cell surface. Similar results were obtained with isolated rat hepatocytes. Moreover, in vesicles of microvillar origin, passive fluxes of Ca2+, ATP, and IP3 occur through cation and anion channels, respectively, suggesting that Ca2+ storage is due to ATP-dependent Ca2+ binding to an intravesicular component. Here we demonstrate that F-actin may be a possible candidate for this function. ATP-actin monomers bind Ca2+ with high affinity (Kd = 2-8 nM) to their divalent cation binding sites. Polymerization of actin monomers decreases the rate constant for divalent cation exchange at this binding site by more than 3 orders of magnitude rendering bound cations nearly unavailable. F-actin-bound Ca2+ can be released by depolymerization and dissociation from Ca(2+)-ADP-actin monomers (Kd = 375 nM). We now provide additional evidence for the possible involvement of actin in Ca2+ storage. (1) Preincubation of surface-derived Ca(2+)-storing vesicles from HIT cells with the F-actin stabilizer, phalloidin, strongly inhibited ATP-dependent Ca2+ uptake, reducing the IP3-sensitive Ca2+ pool by 70%. Phalloidin, when added after the loading process, affected neither the amount of stored Ca2+ nor IP3 action on the store. (2) F-actin polymerized in the presence of Mg2+ in nominally Ca(2+)-free buffer still contained about half of the high affinity sites occupied with Ca2+ (Mg/Ca-F-actin). (3) Using the fura-2 technique, we found that in the presence of ATP, Mg/Ca-F-actin incorporated free Ca2+ at a relatively low rate. Short pulses of ultrasound (3-10 s) strongly accelerated Ca2+ uptake, decreasing free Ca2+ from 500 nM to below 100 nM. (4) In the presence of physiological levels of Mg2+ (0.5 mM), sonication liberated large amounts of Ca2+ from Mg/Ca-F-actin. (5) Ca-F-actin released bound Ca2+ at a very slow rate. Short ultrasonic pulses rapidly elevated free Ca2+ from about 50 nM up to 500 nM. (6) Small amounts of profilin, an actin-binding protein, released Ca2+ both from Ca- and Mg/Ca-F-actin and also inhibited uptake of Ca2+ into Mg/Ca-F-actin. (7) Phalloidin completely inhibited Ca-uptake into Mg/Ca-F-actin even during ultrasonic treatment. These findings suggest that Ca2+ storage may occur by addition of Ca-ATP-actin monomers to reactive ends of the polymer and emptying of this store by profilin-stimulated release of Ca-ADP-actin. Thus, receptor-operated Ca2+ signaling, initiated by phospholipase C activation, may proceed via the well-known phosphatidylinositol phosphate-regulated profilin/gelsolin pathway of actin reorganization/depolymerization. The importance of the proposed microvillar Ca2+ signaling system for living cells remains to be established.
Actins↗
Energy conservation by bifurcated electron-transfer in the cytochrome-bc1 complex.
The overall electron- and proton-pathways within the cytochrome-bc1 complex are described by a widely accepted mechanism known as the protonmotive Q-cycle. Within this reaction scheme, the unique bifurcation of electron flow into a high potential and a low potential pathway occurring at the ubihydroquinone-oxidation center is the energy conserving reaction. It is this reaction, which results in vectorial proton translocation, as it allows the 'recycling' of every second electron across the membrane onto the ubiquinone-reduction center. However, the Q-cycle reaction scheme does not address the detailed chemistry of this central step. Based on a structural model of the ubihydroquinone-oxidation pocket and the assumption that the reaction involves two ubiquinone molecules in a stacked configuration, here I propose a detailed chemical model for the reactions occurring during steady-state catalysis. In this proton-gated charge-transfer mechanism the reaction is controlled by the deprotonation of the substrate ubihydroquinone and not, as proposed earlier, by the formation of a highly unstable semiquinone species.
Animals↗
2-Nitrosofluorene and N-hydroxy-2-aminofluorene react with the ubiquinone-reduction center (center N) of the mitochondrial cytochrome bc1 complex.
We determined the sites of artificial electron transfer onto 2-nitrosofluorene (NOF), a metabolite of carcinogenic 2- acetylaminofluorene in mitochondria and isolated cytochrome bc1 complex. NOF-induced O2 consumption in mitochondria was sensitive to antimycin A, but insensitive to myxothiazol. In the isolated cytochrome bc1 complex, NOF induced rapid MOA-stilbene-insensitive reoxidation of cytochrome b, whereas in the presence of antimycin A, reoxidation was very slow. The corresponding hydroxylamine, N-hydroxy-2-aminofluorene (N-OH-AF), reduced cytochrome b specifically through center N of the cytochrome bc1 complex. We conclude that NOF and N-OH-AF bind to center N of the cytochrome bc1 complex and act as electron acceptor and donor, respectively. The N-OH-AF/NOF interconversion is considered to be involved in the cytotoxicity of 2-acetylaminofluorene in vivo.
Animals↗