Plasma membrane coenzyme Q: evidence for a role in autism.

BACKGROUND: The Voltage Dependent Anion Channel (VDAC) is involved in control of autism. Treatments, including coenzyme Q, have had some success on autism control. DATA SOURCES: Correlation of porin redox activity and expression of autism is based on extensive literature, especially studies of antibodies, identification of cytosolic nicotinamide adenine dinucleotide reduced (NADH) dehydrogenase activity in the VDAC, and evidence for extreme sensitivity of the dehydrogenase to a mercurial. Evidence for a coenzyme Q requirement came from extraction and analog inhibition of NADH ferricyanide reductase in the erythrocyte plasma membrane, done in 1994, and reinterpreted when it was identified in VDAC in 2004. The effects of ubiquinol (the QH2 - reduced form of coenzyme Q) in children with autism were studied. RESULTS: A new role for coenzyme Q in the porin channels has implications on autism. Ubiquinol, the more active form of coenzyme Q, produces favorable response in children with autism. Agents which affected electron transport in porin show parallel effects in autism. CONCLUSION: We propose a hypothesis that autism is controlled by a coenzyme Q-dependent redox system in the porin channels; this conclusion is based on the effects of agents that positively or negatively affect electron transport and the symptoms of autism. The full understanding of the mechanism of their control needs to be established.

Evidence for a relation between plasma membrane coenzyme Q and autism.

Voltage Dependent Anion Channel (VDAC) in the cell membrane transports important molecules and ions across the cell membrane. It was recently shown that VDAC also acts as a trans membrane NADH dehydrogenase. A recent study showed that autistic children have increased antibodies to VDAC proteins and such a binding inhibits both the transport and dehydrogenase activities of VDAC. The derived function of VDAC, therefore, might underlie the development of autism. It has also recently been shown that the dehydrogenase in erythrocyte membranes requires coenzyme Q. Since the plasma membrane oxidase is not in erythrocyte membranes, the coenzyme Q requirement must be for VDAC. This is consistent with sensitivity of the dehydrogenase to SH inhibitors. This is a novel site for coenzyme Q function but it has an analogy with the Q requirement for the mitochondrial uncoupler protein and the permeability transition pore.

Sirtuin activation: a role for plasma membrane in the cell growth puzzle.

For more than 20 years, the observation that impermeable oxidants can stimulate cell growth has not been satisfactorily explained. The discovery of sirtuins provides a logical answer to the puzzle. The NADH-dependent transplasma membrane electron transport system, which is stimulated by growth factors and interventions such as calorie restriction, can transfer electrons to external acceptors and protect against stress-induced apoptosis. We hypothesize that the activation of plasma membrane electron transport contributes to the cytosolic NAD(+) pool required for sirtuin to activate transcription factors necessary for cell growth and survival.

Putting together a plasma membrane NADH oxidase: a tale of three laboratories.

The observation that high cellular concentrations of NADH were associated with low adenylate cyclase activity led to a search for the mechanism of the effect. Since cyclase is in the plasma membrane, we considered the membrane might have a site for NADH action, and that NADH might be oxidized at that site. A test for NADH oxidase showed very low activity, which could be increased by adding growth factors. The plasma membrane oxidase was not inhibited by inhibitors of mitochondrial NADH oxidase such as cyanide, rotenone or antimycin. Stimulation of the plasma membrane oxidase by iso-proterenol or triiodothyronine was different from lack of stimulation in endoplasmic reticulum. After 25 years of research, three components of a trans membrane NADH oxidase have been discovered. Flavoprotein NADH coenzyme Q reductases (NADH cytochrome b reductase) on the inside, coenzyme Q in the middle, and a coenzyme Q oxidase on the outside as a terminal oxidase. The external oxidase segment is a copper protein with unique properties in timekeeping, protein disulfide isomerase and endogenous NADH oxidase activity, which affords a mechanism for control of cell growth by the overall NADH oxidase and the remarkable inhibition of oxidase activity and growth of cancer cells by a wide range of anti-tumor drugs. A second trans plasma membrane electron transport system has been found in voltage dependent anion channel (VDAC), which has NADH ferricyanide reductase activity. This activity must be considered in relation to ferricyanide stimulation of growth and increased VDAC antibodies in patients with autism.

The oxidative function of diferric transferrin.

There is evidence for an unexpected role of diferric transferrin as a terminal oxidase for the transplasma membrane oxidation of cytosolic NADH. In the original studies which showed the reduction of iron in transferrin by the plasma membranes NADH oxidase, the possible role of the reduction on iron uptake was emphasized. The rapid reoxidation of transferrin iron under aerobic conditions precludes a role for surface reduction at neutral pH for release of iron for uptake at the plasma membrane. The stimulation of cytosolic NADH oxidation by diferric transferrin indicates that the transferrin can act as a terminal oxidase for the transplasma membrane NADH oxidase or can bind to a site which activates the oxidase. Since plasma membrane NADH oxidases clearly play a role in cell signaling, the relation of ferric transferrin stimulation of NADH oxidase to cell control should be considered, especially in relation to the growth promotion by transferrin not related to iron uptake. The oxidase can also contribute to control of cytosolic NAD concentration, and thereby can activate sirtuins for control of ageing and growth.

Discovery of plastoquinones: a personal perspective.

The discovery and the rediscovery of plastoquinone (PQ) are described together with the definition of its structure as a 2,3-dimethyl 5 solanosyl benzoquinone. The discovery, by M. Kofler, was a result of a search for Vitamin K. Its rediscovery was made by me, when I was at The Enzyme Institute of the University of Wisconsin, analyzing animals and plants for the newly discovered coenzyme Q. In green plants, I found another lipophilic quinone in addition to coenzyme Q. Some misleading evidence suggested as if the new quinone had coenzyme Q activity in mitochondria, but improved methods gave negative results. When I found that the quinone was concentrated in chloroplasts, I considered a role for it in photosynthesis analogous to the role of coenzyme Q in mitochondria. After moving to the Chemistry Department, University of Texas at Austin, I used a plain light bulb and some spinach chloroplasts to show that PQ could be involved in photosynthetic redox reactions. This effect was supported by Norman Bishop's restoration of chloroplast electron transport after solvent extraction, with PQ and photoreduction studies by E. R. Redfern and J. Friend in R. A. Morton's laboratory in Liverpool, UK. We also found an additional analog of PQ in addition to a second analog found in Wisconsin. We called the new analogs PQB and PQC. Although we found some restoration effects with PQC, the discovery by W. T. Griffiths in Morton's laboratory, that PQB and PQC consisted of six forms of PQ each, made it more likely that the new analogs were breakdown products. Morton's group established the structure of the PQCs as a series of PQs, with a hydroxyl group on the prenyl side chain, and the PQB series as having fatty acids esterified to the hydroxyl groups of PQC. Possible functions of the analogs are also discussed in this article.

Monoascorbate free radical-dependent oxidation-reduction reactions of liver Golgi apparatus membranes.

Golgi apparatus from rat liver contain an ascorbate free radical oxidoreductase that oxidizes NADH at neutral pH with monodehydroascorbate as acceptor to generate a membrane potential. At pH 5.0, the reverse reaction occurs from NAD(+). The electron spin resonance signal of the ascorbate-free radical and its disappearance upon the addition of NADH (pH 7) or NAD(+) (pH 5.0) confirms monodehydroascorbate involvement. Location of monodehydroascorbate both external to and within Golgi apparatus compartments is suggested from energization provided by inward or outward flux of electrons across the Golgi apparatus membranes. The isolated membranes are sealed, oriented cytoplasmic side out and impermeable to NAD(+) and ascorbate. NAD(+) derived through the action of Golgi apparatus beta-NADP phosphohydrolase is simultaneously reduced to NADH with monodehydroascorbate present. The response of the NADH- (NAD(+)-) ascorbate free radical oxidoreductase system to pH in Golgi apparatus provides a simple regulatory mechanism to control vesicle acidification.

The evolution of coenzyme Q.

In the 50 years since the identification of coenzyme Q as an electron carrier in mitochondria, it has been identified with diverse and unexpected functions in cells. Its discovery came as a result of a search for electron carriers in mitochondria following the identification of flavin and cytochromes by Warburg, Keilin, Chance and others. As a result of investigation of membrane lipids at D.E. Green's laboratory at University of Wisconsin coenzyme Q was identified as the electron carrier between primary flavoprotein dehydrogenases and the cytochromes. Then Peter Mitchell identified the role of transmembrane proton transfer as a basis for ATP synthesis. The general distribution of coenzyme Q in all cell membranes then led to the recognition of a role as a primary antioxidant. The protonophoric function was extended to acidification of Golgi and lysosomal vericles. A further role in proton release through the plasma membrane and its relation to cell proliferation has not been fully developed. A role in generation of H202 as a messenger for hormone and cytokine action is indicated as well as prevention of apoptosis by inhibition of ceramide release. Identification of the genes and proteins required for coenzyme Q synthesis has led to a basis for defining deficiency. For 50 years Karl Folkers has led the search for deficiency and therapeutic application. The development of large scale production, better formulation for uptake, and better methods for analysis have furthered this search. The story isn't over yet. Questions remain about effects on membrane structure, breakdown and control of cellular synthesis and uptake and the basis for therapeutic action.

Discovery of ubiquinone (coenzyme Q) and an overview of function.

Details of the discovery of ubiquinone (coenzyme Q) are described in the context of research on mitochondria in the early 1950s. The importance of the research environment created by David E. Green to the recognition of the compound and its role in mitochondria is emphasized as well as the dedicated work of Karl Folkers to find the medical and nutritional significance. The development of diverse functions of the quinone from electron carrier and proton carrier in mitochondria to proton transport in other membranes and uncoupling protein control as well as antioxidant and prooxidant functions is introduced. The successful application in medicine points the way for future development.

Plasma membrane redox and control of sirtuin.

We consider possible contributions of plasma membrane redox systems to Aging control by sirtuin (SIR). Reported changes in plasma membrane redox introduced by calorie restriction (CR) may lead to activation of SIR. The most obvious effect would lie in the increase of NAD+ as a result of NADH oxidation. So the question arises, do the observed changes herald an increase in NADH oxidase under CR? The other possibility is an increase in expression of SIR by activation of plasma membrane oxidase. Previous experiments have shown that activation of the plasma membrane redox system can increase cellular NAD+ concentration. The plasma membrane redox systems are also involved in control of protein kinase activity through oxygen radical generation. This activity may be related to control of SIR expression.

Purification and characterization of a doxorubicin-inhibited NADH-quinone (NADH-ferricyanide) reductase from rat liver plasma membranes.

Plasma membrane-associated redox systems play important roles in regulation of cell growth, internal pH, signal transduction, apoptosis, and defense against pathogens. Stimulation of cell growth and stimulation of the redox system of plasma membranes are correlated. When cell growth is inhibited by antitumor agents such as doxorubicin, capsaicin, and antitumor sulfonylureas, redox activities of the plasma membrane also are inhibited. A doxorubicin-inhibited NADH-quinone reductase was characterized and purified from plasma membranes of rat liver. First, an NADH-cytochrome b(5) reductase, which was doxorubicin-insensitive, was removed from the plasma membranes by the lysosomal protease, cathepsin D. After removal of the NADH-cytochrome b(5) reductase, the plasma membranes retained a doxorubicin-inhibited NADH-quinone reductase activity. The enzyme, with an apparent molecular mass of 57 kDa, was purified 200-fold over the cathepsin D-treated plasma membranes. The purified enzyme had also an NADH-coenzyme Q(0) reductase (NADH: external acceptor (quinone) reductase; EC 1.6.5.) activity. Partial amino acid sequence of the enzyme showed that it was unique with no sequence homology to any known protein. Antibody against the enzyme (peptide sequence) was produced and affinity-purified. The purified antibody immunoprecipitated both the NADH-ferricyanide reductase activity and NADH-coenzyme Q(0) reductase activity of plasma membranes and cross-reacted with human chronic myelogenous leukemia K562 cells and doxorubicin-resistant human chronic myelogenous leukemia K562R cells. Localization by fluorescence microscopy showed that the reaction was with the external surface of the plasma membranes. The doxorubicin-inhibited NADH-quinone reductase may provide a target for the anthracycline antitumor agents and a candidate ferricyanide reductase for plasma membrane electron transport.