The electron transport chain of Bacterionema matruchotii

The membrane fraction of Bacterionema matruchotii contains an electron transport chain with oxidizing activity for NADH and succinate. Respiration was inhibited by KCN, 2-heptyl-4-hydroxyquinoline-N-oxide, UV light irradiation and CO. UV light irradiation, analysis of membrane extracts, and reconstitution of respiration in UV light treated membranes suggested that respiration is mediated by a menaquinone derivative. The membranes contained cytochromes a, b, and c. Inhibition studies and the effect of KCN and CO on the cytochrome spectrum indicated the presence of an a+a3 cytochrome oxidase and cytochrome o. The membrane fraction from cells grown under O2-limiting conditions contained nitrate reductase activity. In B. matruchotii, electron transport is coupled to oxidative phosphorylation as judged by the effects of substrates and inhibitors on the intracellular ATP concentration.     

The effect of changes in the respiratory metabolism upon genome activity in Drosophila

Inhibition of hydrogen transfer between NADH and Co Q by rotenone or amytal in salivary gland cells of Drosophila hydei maintained in vitro, results in the activation of a particular group of four loci in the polytene chromosomes (puff formation). The response of these loci to the same treatment is enhanced if Na-malonate is present in the incubation medium. — Three of the loci become active if the glands are kept in a medium supplied with antimycin A or 2-heptyl-4-hydroxyquinoline-N-oxide (H QNO), specific inhibitors of the electron transfer between cytochromes b and c. — It was established that a temperature treatment and DNP raise oxygen consumption of the cells to a certain level. Following the same treatments of glands supplied with Na-malate and Na-succinate the raise in oxygen consumption attains a significantly higher level. Under these conditions no response is observed at the genome level. — Whereas DNP, which uncouples oxidative phosphorylation and enhances the respiratory chain reactions, does induce the initiation of puff formation, oligomycin, which inhibits oxidative phosphorylation and suppresses the respiratory chain reactions, is ineffective in initiating puff formation at the specific loci. However, if oligomycin is supplied to the medium in combination with KCN which inhibits the cytochrome oxidase activity, three of the four loci become active. — The presence in the medium of substances which may act as hydrogen acceptors, e.g. menadione or methylene blue, can also result in activation of the chromosome loci. — These results are interpreted as indications for the existence of a regulatory mechanism between mitochondrial respiratory metabolism and the activity of a particular group of genome loci.     

Streptolydigin, an Inhibitor of Oxidative Phosphorylation in Rat Liver Mitochondria

Streptolydigin interferes with oxidative phosphorylation in rat liver mitochondria. The agent acts primarily as an uncoupler of respiration-associated phosphorylation but also impairs respiration to various degrees depending on the substrate. Streptolydigin partially inhibits electron flow at a point past the cytochrome b and prior to the cytochrome c reduction site. Streptolydigin also inhibits the function of the enzyme ribonucleic acid polymerase in whole bacterial cells and cell-free systems. The streptolydigin concentrations that cause effective inhibition of ribonucleic acid polymerase in cell-free systems are approximately 10 times less than those required to inhibit oxidative phosphorylation in mitochondria.     

Light-induced redox conversions of cytochrome f in pea leaves]

Redox conversions of cytochrome f were studied in intact pea leaves by double wavelength difference spectrophotometry. Using the inhibition of the photosystem II activity by far red light (719 nm) or diurone, it was found that cytochrome f is located between two photosystems on the oxidative side of photosystem I. The inhibitors of phosphorylation, , e.g. antimycin A and phloridzine, as well as the uncoupler, methylamine, strongly decreased electron transport through the carrier. It is concluded that cytochrome f is functioning in the non-cyclic phosphorylation. It is suggested that in vivo cytochrome f is not coupled with cyclic electron transfer.     

Oxidative Phosphorylation in Fractionated Bacterial Systems: Effect of Chloramphenicol

Chloramphenicol was found to have a direct effect on the respiratory chain of Mycobacterium phlei cells grown in the presence of this drug. Analysis of the respiratory chain components revealed that the presence of chloramphenicol during growth resulted in a partial inhibition in the synthesis of the cytochromes. However, a stimulation in oxidative phosphorylation was observed with the cell-free extract of cells grown in the presence of chloramphenicol. The oxidation of succinate was found to be stimulated 20 to 130%, depending on the particular extract, whereas the oxidation of reduced nicotinamide adenine dinucleotide (NADH) was found to be similar to that of extracts obtained from cells grown in the absence of the drug. Of particular interest was the finding that the cell-free extract of cells grown in the presence of the drug exhibited an increased level of phosphorylation (17 to 100%) when NADH was used as the electron donor. Chloramphenicol appears to affect a component of the respiratory chain between the flavoprotein and cytochrome c. Fractionation of the electron transport particles revealed an increased level of cytochrome b in the fractions which exhibited a stimulation in oxidative phosphorylation.     

Metabolic network control of oxidative phosphorylation: multiple roles of inorganic phosphate

Phosphate (Pi) is a putative cytosolic signaling molecule in the regulation of oxidative phosphorylation. Here, by using a multiparameter monitoring system, we show that Pi controls oxidative phosphorylation in a balanced fashion, modulating both the generation of useful potential energy and the formation of ATP by F1F0-ATPase in heart and skeletal muscle mitochondria. In these studies the effect of Pi was determined on the mitochondria [NADH], NADH generating capacity, matrix pH, membrane potential, oxygen consumption, and cytochrome reduction level. Pi enhanced NADH generation and was obligatory for electron flow under uncoupled conditions. Pi oxidized cytochrome b (cyto-b) and reduced cytochrome c (cyto-c), potentially improving the coupling between the NADH free energy and the proton motive force. The apparent limitation in reducing equivalent flow between cyto-b and cyto-c in the absence of Pi was confirmed in the intact heart by using optical spectroscopic techniques under conditions with low cytosolic [Pi]. These results demonstrate that Pi signaling results in the balanced modulation of oxidative phosphorylation, by influencing both deltaGH+ generation and ATP production, which may contribute to the energy metabolism homeostasis observed in intact systems.     

Defects in the biosynthesis of mitochondrial heme c and heme a in yeast and mammals

Defects in heme biosynthesis have been associated with a large number of diseases, but mostly recognized in porphyrias, which are neurovisceral or cutaneous disorders caused by the accumulation of biosynthetic intermediates. However, defects in the maturation of heme groups that are part of the oxidative phosphorylation system are now also recognized as important causes of disease. The electron transport chain contains heme groups of the types a, b and c, all of which are directly involved in electron transfer reactions. In this article, we review the effect of mutations in enzymes involved in the maturation of heme a (the prosthetic group of cytochrome c oxidase) and heme c (the prosthetic group of cytochrome c) both in yeast and in humans. COX10 and COX15 are two genes, initially identified in Saccharomyces cerevisiae that have been found to cause infantile cytochrome c oxidase deficiency in humans. They participate in the farnesylation and hydroxylation of heme b, steps that are necessary for the formation of heme a, the prosthetic group required for cytochrome oxidase assembly and activity. Deletion of the cytochrome c heme lyase gene in a single allele has also been associated with a human disease, known as Microphthalmia with Linear Skin defects (MLS) syndrome. The cytochrome c heme lyase is necessary to covalently attach the heme group to the apocytochrome c polypeptide. The production of mouse models recapitulating these diseases is providing novel information on the pathogenesis of clinical syndromes.     

Benzoyl peroxide interaction with mitochondria: inhibition of respiration and induction of rapid, large-amplitude swelling

When micromolar concentrations of benzoyl peroxide (BPO) are added to rat liver mitochondria, inhibition of mitochondrial NADH-oxidase and succinoxidase is observed. The addition of 2,4-dinitrophenol, an uncoupler of oxidative phosphorylation, results in only partial release of this inhibition, suggesting that BPO inhibits both electron and energy transfer in mitochondria. Release of inhibition is also observed when an electron donor, N,N,N',N'-tetramethyl-p-phenylenediamine, is added, suggesting that inhibition occurs on the substrate side of cytochrome c. When BPO is added to respiring submitochondrial particles, only reduced cytochrome b is observed to accumulate in the difference spectrum (reduced minus oxidized) in a manner analogous to that observed in the presence of antimycin A. These results indicate that BPO interacts at coupling site II between cytochromes b and c1. When respiring SMP are treated with BPO in the presence of the spin trap 5,5-dimethyl-1-pyrroline-N-oxide, electron spin resonance signals attributable to the hydroxyl and superoxide adducts are observed. Catalase and superoxide dismutase inhibit the formation of these adducts, suggesting the involvement of both hydrogen peroxide and superoxide radicals in this process. BPO also induces rapid, large-amplitude swelling of mitochondria; the swelling is dependent on the presence of monovalent cations but is independent of the presence of calcium, oxygen, and respiratory substrate. BPO-induced swelling appears to be disassociated from radical production and lipid peroxidation.     

Characterisation of Fasciola hepatica cytochrome c peroxidase as an enzyme with potential antioxidant activity in vitro.

Cytochrome c peroxidase oxidises hydrogen peroxide using cytochrome c as the electron donor. This enzyme is found in yeast and bacteria and has been also described in the trematodes Fasciola hepatica and Schistosoma mansoni. Using partially purified cytochrome c peroxidase samples from Fasciola hepatica we evaluated its role as an antioxidant enzyme via the investigation of its ability to protect against oxidative damage to deoxyribose in vitro. A system containing FeIII-EDTA plus ascorbate was used to generate reactive oxygen species superoxide radical, H2O2 as well as the hydroxyl radical. Fasciola hepatica cytochrome c peroxidase effectively protected deoxyribose against oxidative damage in the presence of its substrate cytochrome c. This protection was proportional to the amount of enzyme added and occurred only in the presence of cytochrome c. Due to the low specific activity of the final partially purified sample the effects of ascorbate and calcium chloride on cytochrome c peroxidase were investigated. The activity of the partially purified enzyme was found to increase between 10 and 37% upon reduction with ascorbate. However, incubation of the partially purified enzyme with 1 mM calcium chloride did not have any effect on enzyme activity. Our results showed that Fasciola hepatica CcP can protect deoxyribose from oxidative damage in vitro by blocking the formation of the highly toxic hydroxyl radical (.OH). We suggest that the capacity of CcP to inhibit .OH-formation, by efficiently removing H2O2 from the in vitro oxidative system, may extend the biological role of CcP in response to oxidative stress in Fasciola hepatica.     

Involvement of oxyleghaemoglobin and cytochrome P-450 in an efficient oxidative phosphorylation pathway which supports nitrogen fixation in Rhizobium.

Cellular ATP level, ATP/ADP ratio and nitrogenase activity rise when oxyleghaemoglobin is added to respiring suspensions of Rhizobium japonicum bacteroids from soybean root nodules. Increased gaseous O2 tension is much less efficient than oxyleghaemoglobin in stimulation of bacteroid ATP production. Studies with the inhibitor carbonyl cyanide m-chlorophenylhydrazone show this ATP to be generated as a consequence of oxidative phosphorylation. N-Phenylimidazole, a specific cytochrome P-450 inhibitor, also lowers the efficiency of bacteroid oxidative phosphorylation. An approximately linear relationship is observed between ATP/ADP ratio and nitrogenase activity as N-phenylimidazole concentration is lowered. It is suggested that cytochrome P-450 is a component of the leghaemoglobin-facilitated respiration pathway and that it may act as intracellular O2 carrier rather than terminal oxidase. A less efficient oxidase appears to function when cytochrome P-450 is inhibited.     

Alterations of the Respiratory System of Neurospora crassa by the mi-1 Mutation

The respiratory mechanism of the cytochrome-defective mi-1 mutant of Neurospora crassa was partially characterized and compared with that of the wild type. Both systems of electron transport were linked to oxidative phosphorylation and showed similar characteristics with respect to substrates and components. The system operating in the cytochrome-deficient mi-1 mutant showed no characteristic alpha absorption bands of cytochromes a or b and was very unstable in vitro. The possibility that this system may be an altered form of the normal cytochrome electron transport chain which occurs in the wild type is discussed.     

The effect of cytochrome c and its `dimer'' on electron transfer and energy transformation

A preparation that contained cytochrome c, mainly in the form of its ;dimer', was studied and compared with native cytochrome c with respect to its ability to support electron transfer and energy transformation in cytochrome c-depleted rat liver mitochondria. When the depleted mitochondria were titrated with either cytochrome c or the ;dimer', the extent of coupling between respiration and phosphorylation was enhanced, as manifested by an increase in the P/O ratio. The ;dimer' was relatively ineffective as an electron carrier in the respiratory system, but it was as effective as cytochrome c in reconstitution of oxidative phosphorylation in depleted mitochondria. Addition of ;dimer' to the depleted mitochondria, in the presence of a low, non-saturating concentration of cytochrome c, increased the P/O ratio without concomitant stimulation of respiration. Both cytochrome c and the ;dimer' stimulated spontaneous swelling and electron transport-driven proton translocation in depleted mitochondria. The pattern of action of cytochrome c and its ;dimer' is in accord with the assumption that they affect an early step in energy conservation.     

Roles of residues 129 and 209 in the alteration by cytochrome b5 of hydroxylase activities in mouse 2A P450S.

Cytochrome b5 stimulates the coumarin 7-hydroxylation activity of P450coh. A mutation of Arg-129 in P450coh, however, abolishes the stimulation. Moreover, this mutant P450coh binds loosely to cytochrome b5-conjugated Sepharose 4B, whereas wild-type P450coh binds tightly. Consistent with this, the mutation increases the Ka value for b5 binding approximately 6-fold. The identity of residue 209 also alters the stimulation of the activity of P450coh depending on the type of the substrates used and products formed. Coumarin 7-hydroxylation activity is greatly stimulated by cytochrome b5 only when Phe is at position 209, while cytochrome b5 stimulates testosterone hydroxylation activity of P450coh in which Phe, Asn, Ser or Lys substitutes residue 209. P450coh changes its rate of hydrogen peroxide formation depending on the identity of residue 209 and substrate used. Cytochrome b5 decreases the hydrogen peroxide formation of some P450coh whose activities are stimulated by the cytochrome; however, the decrease does not always result in stimulating the activity. The results indicate, therefore, that residues 129 and 209 play different roles in stimulating P450coh activity by cytochrome b5; Arg-129 is a key residue in the cytochrome b5-binding domain and is essential for the stimulation. Residue 209, however, alters the efficiency of electron transport for substrate oxidation as a residue which resides near the sixth ligand of heme and in the substrate-binding site.     

The mitochondria-targeted imidazole substituted oleic acid ‘TPP-IOA’ affects mitochondrial bioenergetics and its protective efficacy in cells is influenced by cellular dependence on aerobic metabolism

A variety of mitochondria-targeted small molecules have been invented to manipulate mitochondrial redox activities and improve function in certain disease states. 3-Hydroxypropyl-triphenylphosphonium-conjugated imidazole-substituted oleic acid (TPP-IOA) was developed as a specific inhibitor of cytochrome c peroxidase activity that inhibits apoptosis by preventing cardiolipin oxidation and cytochrome c release to the cytosol. Here we evaluate the effects of TPP-IOA on oxidative phosphorylation in isolated mitochondria and on mitochondrial function in live cells. We demonstrate that, at concentrations similar to those required to achieve inhibition of cytochrome c peroxidase activity, TPP-IOA perturbs oxidative phosphorylation in isolated mitochondria. In live SH-SY5Y cells, TPP-IOA partially collapsed mitochondrial membrane potential, caused extensive fragmentation of the mitochondrial network, and decreased apparent mitochondrial abundance within 3 h of exposure. Many cultured cell lines rely primarily on aerobic glycolysis, potentially making them less sensitive to small molecules disrupting oxidative phosphorylation. We therefore determined the anti-apoptotic efficacy of TPP-IOA in SH-SY5Y cells growing in glucose or in galactose, the latter of which increases reliance on oxidative phosphorylation for ATP supply. The anti-apoptotic activity of TPP-IOA that was observed in glucose media was not seen in galactose media. It therefore appears that, at concentrations required to inhibit cytochrome c peroxidase activity, TPP-IOA perturbs oxidative phosphorylation. In light of these data it is predicted that potential future therapeutic applications of TPP-IOA will be restricted to highly glycolytic cell types with limited reliance on oxidative phosphorylation.     

The role of electron transport in the defence response of the South African abalone, Haliotis midae

In order to establish health management systems for farmed abalone, it is necessary to understand how the abalone immune system functions and responds to stimulation. Two electron transport system genes, cytochrome b and cytochrome c oxidase III, were found to be upregulated in a cDNA microarray experiment performed on haemocytes from immune-stimulated abalone (Arendze-Bailey, unpublished). The current study sought to elucidate the role of these genes, and thus the electron transport system, in the abalone immune response by specifically inhibiting cytochrome b with antimycin A and measuring haemocyte immune parameters in vivo. Antimycin A did not decrease haemocyte cell viability, but halved cellular ATP from 4 x 10(12) nM/cell to 2 x 10(12) nM/cell (p < 0.05, unpaired t-test). Inhibition of electron transport resulted in a 0.6 fold increase in cellular superoxide levels (p < 0.05, unpaired t-test), while phagocytosis dropped by nearly 50% (p < 0.05, ANOVA) and the ability of haemocytes to kill bacteria was also reduced. Since cytochrome b and cytochrome c oxidase III expression is upregulated in immune-stimulated abalone, and inhibition of electron transport resulted in a decreased immune response in vivo, we conclude that the abalone immune response is dependent on electron transport and that oxidative phosphorylation plays a role in the immune response following stimulation.     

Cytochrome c-catalyzed oxidation of organic molecules by hydrogen peroxide.

Cytochrome c catalyzed the oxidation of various electron donors in the presence of hydrogen peroxide (H2O2), including 2-2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 4-aminoantipyrine (4-AP), and luminol. With ferrocytochrome c, oxidation reactions were preceded by a lag phase corresponding to the H2O2-mediated oxidation of cytochrome c to the ferric state; no lag phase was observed with ferricytochrome c. However, brief preincubation of ferricytochrome c with H2O2 increased its catalytic activity prior to progressive inactivation and degradation. Superoxide (O2-) and hydroxyl radical (.OH) were not involved in this catalytic activity, since it was not sensitive to superoxide dismutase (SOD) or mannitol. Free iron released from the heme did not play a role in the oxidative reactions as concluded from the lack of effect of diethylenetriaminepentaacetic acid. Uric acid and tryptophan inhibited the oxidation of ABTS, stimulation of luminol chemiluminescence, and inactivation of cytochrome c. Our results are consistent with an initial activation of cytochrome c by H2O2 to a catalytically more active species in which a high oxidation state of an oxo-heme complex mediates the oxidative reactions. The lack of SOD effect on cytochrome c-catalyzed, H2O2-dependent luminol chemiluminescence supports a mechanism of chemiexcitation whereby a luminol endoperoxide is formed by direct reaction of H2O2 with an oxidized luminol molecule, either luminol radical or luminol diazoquinone.     

Effect of organophosphorus insecticides on the oxidative processes in rat brain synaptosomes

Abstract: This paper describes the effect of four organophosphorus insecticides: Dipterex, DDVP, Ronnel and its oxygen analogue on the respiration of rat brain synaptosomes. Dipterex and DDVP in the concentrations used, 5, 50, or 500 μM, did not change the rate of oxygen uptake and oxidative phosphorylation in rat brain synaptosomes. Ronnel in the highest concentration (500 μM) inhibited respiration in state 3 conditions and abolished respiratory control by ADP. This inhibition was correlated with a change of cytochrome c oxidase activity. The oxygen analogue of Ronnel (OAR) in micromolar concentrations (50 μM) increased the rate of respiration of synaptosomes utilizing glutamate plus malate as substrate. Higher concentrations of OAR produced inhibition of respiration, cytochrome c oxidase and NADH: cytochrome c reductase activities. These observations are typical for uncouplers of oxidative phosphorylation. Noteworthy is the fact that the uncoupling activity of OAR was observed at concentrations which did not inhibit acetylcholinesterase activity. These findings seem to suggest that disturbances in oxidative processes could play an important role in the toxicity of organophosphorus insecticides. The relation between chemical structure and the ability of insecticides to affect oxidative phosphorylation is discussed.     

Influence of nitrate on fermentation pattern, molar growth yields and synthesis of cytochrome b in Propionibacterium pentosaceum.

Under anaerobic conditions, Propionibacterium pentosaceum reduces nitrate to nitrite until nitrate is exhausted from the medium when nitrite is converted into N2 or N2O. In the presence of nitrate, fermentation patterns for lactate, glycerol and pyruvate were different from those obtained during anaerobic growth without an inorganic electron acceptor. In the presence of these substrates, a drastic decrease in propionate formation was observed, some pyruvate accumulated during growth with lactate, and acetate was produced from glycerol. Acetate production from lactate and pyruvate was not influenced by the presence of nitrate. Furthermore, CO2 was produced by citric acid cycle activity. The fermentation pattern during nitrite reduction resembled that of P. pentosaceum grown anaerobically without an inorganic electron acceptor. Nitrits has a toxic effect, since bacteria inoculated into a medium with 9 mM-nitrite failed to grow. The cytochrome spectrum of anaerobically grown P. pentosaceum was similar with and without nitrate. In membrane fractions of bacteria grown anaerobically with nitrate, cytochrome b functioned in the transfer of electrons from lactate, glycerol I-phosphate and NADH to nitrate. Molar growth yeilds were increased in the presence of nitrate, indicating an increased production of ATP. This could be explained by citric acid cycle activity, and by ocidative phosphorylation coupled to nitrate reduction. Assuming that I mol ATP is formed in the electron transfer from lactate or glycerol I-phosphate to nitrate, and that 2 mol ATP are formed in the electron transfer from NADH to nitrate, YATP values (g dry wt bacteria/mol ATP) were obtained of between 5-0 and 12-6. The higher YATP values were similar to those obtained during anaerobic growth without an inorganic electron acceptor. This supports the assumptions about the efficiency of oxidative phosphorylation for electron transport to nitrate. Low YAPT values were found when high concentrations of nitrite (15 to 50 mM) accumulated, and were probably due to the toxic effect of nitrite.     

P450 reductase and cytochrome b5 interactions with cytochrome P450: effects on house fly CYP6A1 catalysis

The interactions of protein components of the xenobiotic-metabolizing cytochrome P450 system, CYP6A1, P450 reductase, and cytochrome b5 from the house fly (Musca domestica) have been characterized. CYP6A1 activity is determined by the concentration of the CYP6A1-P450 reductase complex, regardless of which protein is present in excess. Both holo- and apo-b5 stimulated CYP6A1 heptachlor epoxidase and steroid hydroxylase activities and influenced the regioselectivity of testosterone hydroxylation. The conversion of CYP6A1 to its P420 form was decreased by the addition of apo-b5. The effects of cytochrome b5 may involve allosteric modification of the P450 enzyme that modify the conformation of the active site. The overall stoichiometry of the P450 reaction was substrate-dependent. High uncoupling of CYP6A1 was observed with generation of hydrogen peroxide, in excess over the concomitant testosterone hydroxylation or heptachlor epoxidation. Inclusion of cytochrome b5 in the reconstituted system improved efficiency of oxygen consumption and electron utilization from NADPH, or coupling of the P450 reaction. Depending on the reconstitution conditions, coupling efficiency varied from 8 to 25% for heptachlor epoxidation, and from 11 to 70% for testosterone hydroxylation. Because CYP6A1 is a P450 involved in insecticide resistance, this suggests that xenobiotic metabolism by constitutively overexpressed P450s may be linked to significant oxidative stress in the cell that may carry a fitness cost.     

Role of cytochrome c as a stimulator of alpha-synuclein aggregation in Lewy body disease.

alpha-Synuclein is a major component of aggregates forming amyloid-like fibrils in diseases with Lewy bodies and other neurodegenerative disorders, yet the mechanism by which alpha-synuclein is intracellularly aggregated during neurodegeneration is poorly understood. Recent studies suggest that oxidative stress reactions might contribute to abnormal aggregation of this molecule. In this context, the main objective of the present study was to determine the potential role of the heme protein cytochrome c in alpha-synuclein aggregation. When recombinant alpha-synuclein was coincubated with cytochrome c/hydrogen peroxide, alpha-synuclein was concomitantly induced to be aggregated. This process was blocked by antioxidant agents such as N-acetyl-L-cysteine. Hemin/hydrogen peroxide similarly induced aggregation of alpha-synuclein, and both cytochrome c/hydrogen peroxide- and hemin/hydrogen peroxide-induced aggregation of alpha-synuclein was partially inhibited by treatment with iron chelator deferoxisamine. This indicates that iron-catalyzed oxidative reaction mediated by cytochrome c/hydrogen peroxide might be critically involved in promoting alpha-synuclein aggregation. Furthermore, double labeling studies for cytochrome c/alpha-synuclein showed that they were colocalized in Lewy bodies of patients with Parkinson's disease. Taken together, these results suggest that cytochrome c, a well known electron transfer, and mediator of apoptotic cell death may be involved in the oxidative stress-induced aggregation of alpha-synuclein in Parkinson's disease and related disorders.