Regulation and termination of NADPH oxidase activity

NADPH oxidase of phagocytes plays a crucial role in host defense by producing reactive oxygen species (ROS) that are intended to kill invading microbes. Many other cells produce ROS for signaling purposes. The respiratory burst oxidase in human neutrophils is the main but not exclusive subject of this review, because it is archetypical and has been studied most extensively. The activity of this enzyme must be controlled in phagocytes to prevent collateral damage, and in non-phagocytic cells to perform its signaling role. With many stimuli, NADPH oxidase activity is transient. Various forms of evidence indicate that sustained NADPH oxidase activity requires continuous renewal of the enzyme complex, without which rapid deactivation occurs. This review considers mechanisms that have been proposed to terminate the phagocyte respiratory burst. Changes in the phosphorylation state of p47(phox) and in the species of nucleotide bound to Rac seem to be the dominant factors in deactivation.     

The superoxide-generating NADPH oxidase: structural aspects and activation mechanism

Flavocytochrome b ₅₅₈ is the catalytic core of the respiratory-burst oxidase, an enzyme complex that catalyzes the NADPH-dependent reduction of O₂ into the superoxide anion O₂ ^- in phagocytic cells. Flavocytochrome b ₅₅₈ is anchored in the plasma membrane. It is a heterodimer that consists of a large glycoprotein gp91phox (phox for phagocyte oxidase) (β subunit) and a small protein p22phox (α subunit). The other components of the respiratory-burst oxidase are water-soluble proteins of cytosolic origin, namely p67phox, p47phox, p40phox and Rac. Upon cell stimulation, they assemble with the membrane-bound flavocytochrome b ₅₅₈ which becomes activated and generates O₂ ^-. A defect in any of the genes encoding gp91phox, p22phox, p67phox or p47phox results in chronic granulomatous disease, a genetic disorder characterized by severe and recurrent infections, illustrating the role of O₂ ^- and the derived metabolites H₂O₂ and HOCl in host defense against invading microorganisms. The electron carriers, FAD and hemes b, and the binding site for NADPH are confined to the gp91phox subunit of flavocytochrome b ₅₅₈ . The p22phox subunit serves as a docking site for the cytosolic phox proteins. This review provides an overview of current knowledge on the structural organization of the O₂ ^--generating flavocytochrome b ₅₅₈ , its kinetics, its mechanism of activation and the regulation of its biosynthesis. Homologues of gp91phox, called Nox and Duox, are present in a large variety of non-phagocytic cells. They exhibit modest O₂ ^--generating oxidase activity, and some act as proton channels. Their role in various aspects of signal transduction is currently under investigation and is briefly discussed. Received 28 May 2002; received after revision 20 June 2002; accepted 24 June 2002     

Potentiation of histamine and inhibition of diamine oxidase by mescaline

Zusammenfassung Anhand zweier Versuchsanordnungen wird gezeigt, dass Meskalin Histaminwirkungen verstärken kann. Ausserdem vermag es die Histamin zerstörende Wirkung der Diaminooxydase zu hemmen (in 10^–5-molarer Konzentration).Es wird die Möglichkeit diskutiert, dass diese Eigenschaften mit der 4-Methoxyphenyl-äthylamin-Struktur des Meskalins in Zusammenhang stehen könnten.     

Monoamine oxidase inhibition by d-amphetamine in ganglia and nerve endings

Summary The inhibition of monoamine oxidase (MAO) activity by d-amphetamine was measured in homogenates of cat superior cervical ganglion and nictitating membrane, using tyramine (TM) and noradrenaline (NA) as substrates. In both tissues, d-amphetamine was shown to be a competitive inhibitor of the oxidation of TM. The Ki for d-amphetamine, as a MAO inhibitor, was lower in the ganglia than in the peripheral nerve endings.Supported by a Contract from the National Research Council of Argentina (CONICET) (Res. 67/79).     

Oxidation of vanillin by cream xanthine oxidase and its inhibition by allopurinol

Zusammenfassung Es wird gezeigt, dass Vanillin durch Xanthinoxydase oxydiert und dass diese Reaktion durch Allopurinol gehemmt wird.     

Lack of specific inhibition of graft-versus-host reaction by anti-fab serum

Résumé En recherchant une action des anticorps antiFab sur la réaction primaire des cellules immunologiquement compétentes, on a mis en évidence une inhibition de la réaction du greffon contre l'hôte par du sérum de Lapin anti-Fab de Souris: cette inhibition n'est cependant pas liée aux anticorps anti-Fab, mais à la cytotoxicité des sérums de Lapin aussi bien anti-Fab que normal.     

The tryptophans in flavodoxin and synthetic flavinyl peptides characterized by chemical and photochemical oxidations

Zusammenfassung Zwei der vier Tryptophan-Reste in Flavodoxin sind relativ gut abgeschirmt, wie das Ausmass der Oxidation durch N-Bromsuccinimid zeigt. Ausserdem kann wahrscheinlich mindestens einer dieser Tryptophan-Reste mit FMN einen stabilen Komplex bilden, wie die Unempfindlichkeit gegenüber Photooxidation nahelegt.     

Antimicrobial peptides: natural templates for synthetic membrane-active compounds

The innate immunity of multicellular organisms relies in large part on the action of antimicrobial peptides (AMPs) to resist microbial invasion. Crafted by evolution into an extremely diversified array of sequences and folds, AMPs do share a common amphiphilic 3-D arrangement. This feature is directly linked with a common mechanism of action that predominantly (although not exclusively) develops upon interaction of peptides with cell membranes of target cells. This minireview reports on current understanding of the modes of interaction of AMPs with biological and model membranes, especially focusing on recent insights into the folding and oligomerization requirements of peptides to bind and insert into lipid membranes and exert their antibiotic effects. Given the potential of AMPs to be developed into a new class of anti-infective agents, emphasis is placed on how the information on peptide-membrane interactions could direct the design and selection of improved biomimetic synthetic peptides with antibiotic properties.     

NADPH oxidase inhibitors: a decade of discovery from Nox2ds to HTS

NADPH oxidases (Nox) are established as major sources of reactive oxygen species (ROS). Over the past two decades, Nox-derived ROS have emerged as pivotal in the development of myriad diseases involving oxidative stress. In contrast, Nox are also involved in signaling mechanisms necessary for normal cell function. The study of these enzymes in physiological and pathophysiological conditions is made considerably more complex by the discovery of 7 isoforms: Nox1 through 5 as well as Duox1 and 2, each with its own specific cytosolic components, regulatory control mechanisms, subcellular localization and/or tissue distribution. A clear understanding of the role individual isoforms play in a given system is hindered by the lack of isoform-specific inhibitors. In animal models, knockdown or knockout methodologies are providing definitive answers to perplexing questions of the complex interplay of multiple Nox isoforms in cell and tissue signaling. However, the complex structures and interactions of these heteromeric isozymes predict pleiotropic actions of the Nox subunits and thus suppression of these proteins is almost certain to have untoward effects. Thus, as both therapies and pharmacological tools, molecule-based inhibitors continue to prove extremely useful and rational in design. Unfortunately, many of the available inhibitors have proven non-specific, falling into the category of scavengers or inhibitors of more than one source of ROS. Here, we will review some of the efforts that have been undertaken to develop specific inhibitors of NADPH oxidase over the past decade, from the peptidic inhibitor Nox2ds-tat to more recent small molecule inhibitors that have emerged from high-throughput screening campaigns.     

Base pairing in messenger RNA's for small peptides

Summary Longest runs of Watson-Crick pairing in hypothetical m-RNA's for a number of natural peptides were no greater than those in the hypothetical m-RNA's for a large number of randomized amino acid sequences from these peptides. This shows that even if base-pairing in m-RNA were a biological requirement, it would little constrain the amino acid sequence.     

Base pairing in messenger RNA's for small peptides

Longest runs of Watson-Crick pairing in hypothetical m-RNA's for a number of natural peptides were no greater than those in the hypothetical m-RNA's for a large number of randomized amino acid sequences from these peptides. This shown that even if base-pairing in m-RNA were a biological requirement, it would little constrain the amino acid sequence.     

Cadmium-induced inhibition of brain monoamine oxidase in the freshwater catfish

Summary The freshwater catfishClarias batrachus (L.), exposed to 50 ppm of cadmium (Cd) chloride for a period of 135 days exhibited marked inhibition of brain monoamine oxidase (MAO) (p<0.001). The retarded gonadal growth observed in the present study suggests that Cd may be capable of creating imbalance in the aminergic activity of the hypothalamus, which modulates the secretion and release of gonadotropins.Acknowledgment. Financial assistance from the Council of Scientific and Industrial Research of India made this work possible.     

On the specific inhibition of adrenal steroid biosynthesis

Zusammenfassung Der Einfluss von Su-4885 (Metopiron®), Su-8000, Su-9055 und Su-10'603 auf die Corticosteroid-Biosynthese wurdein vitro und teilsin vivo untersucht. Ausser den bekannten Hemmeffekten auf die 11- und 17-Hydroxylierung wurden davon unabhängig auch solche auf die Bildung von Aldosteron, 18-Hydroxy- und 19-Hydroxycorticosteron beobachtet.     

Potentiation of the inhibition of xanthine oxidase by a combination of 6-mercaptopurine and 6-thioguanine

Summary Carcinostatic agents, 6-mercaptopurine and 6-thioguanine inhibited the in vitro and in vivo activity of the enzyme xanthine oxidase (xanthine-oxygen oxidoreductase E.C. 1.2.3.2.). Simultaneous, addition of a mixture of the 2 antimetabolites produced a synergistic effect on the inhibition of the enzyme activity.We thank the University Grants Commission, New Delhi, for their financial assistance and the Dean, M.G.M. Medical College Indore, for providing necessary facilities.     

Regulation of lactate dehydrogenase activity: reversible and isoenzyme-specific inhibition of the tetramerization process by peptides

The transition of inactive lactic dehydrogenase (LDH) subunits to functional tetramers is controlled by 2 naturally-occurring peptides. One of these peptides inhibits the folding/association/activation process of H-LDH and the other peptide, the reconstitution of M-LDH, NADH and NAD+ are isoenzyme-specific antagonists of the 2 inhibitory peptides.     

Depressor effect of synthetic peptides related to ACTH on blood pressure in rats

Summary The depressor effects of natural and synthetic ACTH peptides were demonstrated in the rat. This is an extra-adrenal action of ACTH and is not related to the adrenal-stimulating or melanocyte-stimulating activity of the peptide.