Copper-containing amine oxidases. Biogenesis and catalysis; a structural perspective

This review will focus on how X-ray crystallographic studies of copper-containing amine oxidases have complemented the solution, kinetic, and spectroscopic research on this ubiquitous class of enzymes. These enzymes not only contain a copper ion at the active site, but also a unique organic cofactor, 2,4,5-trihydroxyphenylalanine quinone (TPQ), which is absolutely required for catalysis. Structural data have not only shed light on the catalytic mechanism of the enzyme, which converts primary amines, using molecular oxygen, to aldehydes, ammonia, and hydrogen peroxide, but also on biogenesis of the cofactor. The cofactor is derived from a tyrosine in the enzyme amino acid sequence and requires only the addition of copper(II) and molecular oxygen in a self-processing event.     

Thirty years of microbial P450 monooxygenase research: peroxo-heme intermediates--the central bus station in heme oxygenase catalysis

Oxygen has always been recognized as an essential element of many life forms, initially through its role as a terminal electron acceptor for the energy-generating pathways of oxidative phosphorylation. In 1955, Hayaishi et al. [Mechanism of the pyrocatechase reaction, J. Am. Chem. Soc. 77 (1955) 5450-5451] presented the most important discovery that changed this simplistic view of how Nature uses atmospheric dioxygen. His discovery, the naming and mechanistic understanding of the first "oxygenase" enzyme, has provided a wonderful opportunity and scientific impetus for four decades of researchers. This volume provides an opportunity to recognize the breakthroughs of the "Hayaishi School." Notable have been the prolific contributions of Professor Ishimura et al. [Oxygen and life. Oxygenases, Oxidases and Lipid Mediators, International Congress Series, Elsevier, Amsterdam, 2002], a first-generation Hayaishi product, to characterization of the cytochrome P450 monooxygenases.     

The signaling pathway of NADPH oxidase and its role in glomerular diseases

Abstract

Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (Nox), a major source of reactive oxygen species, is a critical mediator of redox signaling. It is well-documented that oxidative stress is associated with the development of glomerular diseases (GN). Hence, the Nox was also thought to be involved in the pathogenesis of GN. However, the expression of Nox in various GN was not consistent, the mechanisms by which the activity of the Nox enzymes in regulating renal cells remains unclear. Signaling pathways might be very important in the pathogenesis of GN. We performed this review to provide a relatively complete signaling pathways flowchart for Nox to the investigators who were interested in the role of Nox in the pathogenesis of GN. Here, we reviewed the signal transduction pathway of Nox and its role in the pathogenesis of GN.     

Oxygen reactions with bacterial oxidases and globins: binding,reduction and regulation

Oxygen is favoured as terminal electron acceptor in aerobic and facultative microorganisms because of its appropriate physical state, satisfactory solubility and its desirable combinations of kinetic and thermodynamic properties. Oxygen is generally reduced by four electrons to yield oxygen, but there are important biological consequences of, and roles for, the partial reduction to superoxide and peroxide. Complex and multiple regulatory networks ensure (i) the utilization of oxygen in preference to other oxidants, (ii) the synthesis of oxygen-consuming enzymes with appropriate properties (particularly affinity for the ligand), and (iii) appropriate cellular protection in the event of oxidative stress. This contribution reviews the terminal respiratory oxidases of selected Gram-negative bacteria and microbial haemoglobin-like proteins.Recent studies of the cytochromebd-type oxidases ofEscherichia coli andAzotobacter vinelandii suggest that, despite probable similarity at the amino acid level, the reactivities of these oxidases with oxygen are strikingly different. The respiratory protection afforded to nitrogenase in the obligately aerobic diazotrophA. vinelandii by the cytochromebd complex appears to be accompanied by, and may be the result of, a low affinity for oxygen and a high V_max. The poorly characterized cytochromeo-containing oxidase in this bacterium is not required for respiratory protection. InE. coli, the cytochromebd-type oxidase has a remarkably high affinity for oxygen, consistent with the view that this is an oxygen-scavenging oxidase utilized under microaerobic conditions. The demonstration of substrate (i.e. oxygen) inhibition in this complex suggests a mechanism whereby wasteful electron flux through a non-proton-pumping oxidase is avoided at higher dissolved oxygen tensions. The demonstration of two ligandbinding sites (haemsd andb ₅₉₅) in oxidases of this type suggests plausible mechanisms for this phenomenon. InE. coli, assembly of the cytochromebd-type oxidase (and of periplasmic cytochromesb andc) requires the presence of an ABC transporter, which may serve to export haem or some assembly factor' to the periplasm.There is at least one additional oxygen-consuming protein inE. coli — the flavohaemoglobin encoded by thehmp gene. Globin-like proteins are also widely distributed in other bacteria, fungi and protozoa, but most have unknown functions. The function of HMP and the related chimaeric flavohaemoglobins in other bacteria and yeast is unknown; one of several possibilities for HMP is that its relatively low affinity for oxygen during turnover with NADH as substrate could enable it to function as a sensor of falling (or rising) cytoplasmic oxygen concentrations.(until October 1994: Section of Microbiology, Wing Hall, Cornell University, Ithaca, NY 14853-8101, USA)     

Innovation and tinkering in the evolution of oxidases

Although molecular oxygen is a relative newcomer to the biosphere, it has had a profound impact on metabolism. About 700 oxygen‐dependent enzymatic reactions are known, the vast majority of which emerged only after the appearance of oxygen in the biosphere, circa 3 billion years ago. Oxygen was a major driving force for evolutionary innovation—~60% of all known oxygen‐dependent enzyme families emerged as such; that is, the founding ancestor was an O₂‐dependent enzyme. The other 40% seem to have diverged by tinkering from pre‐existing proteins whose function was not related to oxygen. Here, we focus on the latter. We describe transitions from various enzyme classes, as well as from non‐enzymatic proteins, and we explore these transitions in terms of catalytic chemistry, metabolism, and protein structure. These transitions vary from subtle ones, such as simply repurposing oxidoreductases by replacing an electron acceptor such as NAD by O₂, to drastic changes in reaction mechanism, such as turning carboxylases and hydrolases into oxidases. The latter is more common and can occur with strikingly minor changes, for example, only one mutation in the active site. We further suggest that engineering enzymes to harness the extraordinary reactivity of oxygen may yield higher catabolic power and versatility.     

Jonathon Howard.

Jonathon 'Joe' Howard (Fig. 1) is Group Leader and Director at the Max Planck Institute of Molecular Cell Biology and Genetics; he and his research group moved to Dresden, Germany, in July 2001. Howard received his PhD in neurobiology in 1983 from the Australian National University in Canberra. He did postdoctoral research there and also at the University of Bristol, UK, and at the University of California, San Francisco. In 1989, he joined the faculty at the University of Washington. His book "Mechanics of Motor Proteins and the Cytoskeleton" was published earlier this year. [interview by Mari N. Jensen]     

A Possible Evolutionary Origin for the Mn<Subscript>4</Subscript> Cluster in Photosystem II: From Manganese Superoxide Dismutase to Oxygen Evolving Complex

The recently published X-ray absorption fine structure of photosystem II provides a more detailed architecture of the oxygen-evolving complex (OEC) and the surrounding amino acids. In this paper, a comparison between manganese superoxide dismutase, dinuclear manganese catalase enzymes and the oxygen evolving complex in photosystem II is reported. The author suggests that the development of oxygenic photosynthesis occurred in steps, the first of which involved only one manganese ion (Mn(II)) that oxidized two water molecules to hydrogen peroxide and then oxygen.     

Howard A. Schneiderman--Planetary Patriot

SYNOPSIS. This symposium on arthropod development is dedicatedto the memory of Howard A. Schneiderman, who died on December5, 1990. Howard devoted much of his professional life to researchon arthropods and was the author of numerous publications onthe developmental biology of moths and flies. At Case-WesternReserve University and at the University of California-Irvinehe founded research institutes devoted to developmental biologyon arthropods as well as other organisms. Howard Schneidermanchampioned the use of insect growth regulators, which derivedfrom his research on the chemistry and physiology of juvenilehormone, as environmentally innocuous methods of controllinginsect pests of our food and fiber. In recent years he was aproponent of the use of molecular biology to alter the plantgenome so that insect-resistant crops might be grown and therebyreduce the use of insecticides. His global perspective was evidentin his understanding of how biotechnology could be applied toworld agriculture. Moreover, he quickly achieved prominencein promoting industry-university relations in his capacity asSenior Vice President for Research and Development at the MonsantoCompany. The cooperation among academic and industrial institutionsthat he fostered stands as a model for such relationships. HowardSchneiderman's influence on science and its applications, andon the universityindustry interface was profound and will befelt for many years to come.     

Cleavage of pyrogallol by non-heme iron-containing dioxygenases

Both intradiol and proximal extradiol dioxygenases are thought to produce the same product, alpha-hydroxymuconic acid, when pyrogallol (3-hydroxycatechol) is used as a substrate. However, when these enzymes were reacted with pyrogallol, they gave different products. A proximal extradiol dioxygenase, metapyrocatechase (catechol:oxygen 2,3-d-oxidoreductase (decyclizing), EC 1.13.11.2), gave a product having an absorption maximum at 290 nm, which was gradually converted to a more stable compound having an absorption maximum at 239 nm. On the other hand, an intradiol dioxygenase, protocatechuate 3,4-dioxygenase (protocatechuate:oxygen 3,4-oxidoreductase (decyclizing), EC 1.13.11.3), gave a product having an absorption maximum at 300 nm. Based on the spectral data and direct comparison with authentic samples, the primary products obtained by the action of the former and the latter enzymes were identified as alpha-hydroxymuconic acid and 2-pyrone-6-carboxylic acid, respectively. While another intradiol dioxygenase, pyrocatechase (catechol:oxygen 1,2-oxidoreductase (decyclizing), EC 1.13.11.1), gave a mixture of nearly equimolar amounts of these two compounds. Isotope labeling experiments indicated that 1 atom of oxygen was incorporated in 2-pyrone-6-carboxylic acid from the atmosphere. Based on these findings, the reaction mechanism for the formation of 2-pyrone-6-carboxylic acid is discussed. This may be the first experimental evidence indicating the presence of a seven-membered lactone intermediate during the oxygenative cleavage of catechols, proposed by Hamilton (Hamilton, G.A. (1974) in Molecular Mechanisms of Oxygen Activation (Hayaishi, O., ed) pp. 405-451, Academic Press, New York).     

Upstream Regulators and Downstream Effectors of NADPH Oxidases as Novel Therapeutic Targets for Diabetic Kidney Disease

Oxidative stress has been linked to the pathogenesis of diabetic nephropathy, the complication of diabetes in the kidney. NADPH oxidases of the Nox family, and in particular the homologue Nox4, are a major source of reactive oxygen species in the diabetic kidney and are critical mediators of redox signaling in glomerular and tubulointerstitial cells exposed to the diabetic milieu. Here, we present an overview of the current knowledge related to the understanding of the role of Nox enzymes in the processes that control mesangial cell, podocyte and tubulointerstitial cell injury induced by hyperglycemia and other predominant factors enhanced in the diabetic milieu, including the renin-angiotensin system and transforming growth factor-β. The nature of the upstream modulators of Nox enzymes as well as the downstream targets of the Nox NADPH oxidases implicated in the propagation of the redox processes that alter renal biology in diabetes will be highlighted.     

Confusion and controversy in the stress field.

An attempt is made to further clarify present areas of controversy in the stress field, in response to a two-part article by Dr. John W. Mason which concludes in this issue of the Journal of Human Stress. The author tries to elucidate each source of confusion enumerated by Dr. Mason. The continued use of the word "stress" for the nonspecific response to any demand is deemed most desirable. The once vague term can now be applied in a well-defined sense and is accepted in all foreign languages as well, including those in which no such word existed previously in any sense. Subdivision of the stress concept has become necessary as more recent work has led to such notions as "eustress," "distress," "systemic stress" and "local stress." Confusion between stress as both an agent and a result can be avoided only by the distinction between "stress" and "stressor". It is explained that the stress syndrome is--by definition--nonspecific in its causation. However, depending upon conditioning factors, which can selectively influence the reactivity of certain organs, the same stressor can elicit different manifestations in different individuals.     

How does the cold stenothermal gadoid<Emphasis Type="Italic"> Lota lota</Emphasis> survive high water temperatures during summer?

The cold-stenothermal freshwater gadid Lota lota inhabiting the potamic regions of lowland rivers in central Europe, is exposed to summer temperatures up to 25 °C, which is far above the thermal preferendum of this species. Oxygen consumption rates, determined in field catches sampled at different times of the year, revealed that the basal metabolic rate is depressed during summer when water temperatures are high (152±16 mol O₂ 100 g^–1 h^–1at 22 °C in July compared to 250±33 mol O₂ 100 g^–1 h^–1 at 6 °C in November). This observation led us to investigate whether the observed depression of the metabolic rate is caused by oxygen limitation due to thermal impairment of the ventilatory system, as has been observed in other species. Determination of anaerobic end products (lactate and succinate) in the liver tissue of fish caught at different sampling dates did not show an accumulation of anaerobic end products during the summer, indicating no oxygen limitation. Measurements of enzyme activities in the white musculature and liver suggest that enzymes involved in aerobic metabolism were down-regulated during summer, which may have contributed to the observed reduction of metabolic rate.Abbreviations CS citric synthase - LDH lactate dehydrogenase - PK pyruvate kinase - TCA trichloroacetic acid Communicated by G. Heldmaier     

Properties of the stable aerobic and anaerobic half-reduced states of NADPH-cytochrome c reductase.

The microsomal flavoprotein, NADPH-cytochrome c reductase, has been reexamined to determine: (1) the nature of the flavine bound to the enzyme and (2) the oxidation-reduction state of the "half-reduced" form of the flavoprotein. Iyanagi and Mason (Iyanagi, T., and Mason, H.S. (1973), Biochemistry 12, 2297) have recently proposed that NADPH-cytochrome c reductase contains both FAD and FMN as prosthetic groups in lieu of FAD as the sole constituent, as suggested by all previous studies of this enzyme. The data presented herein, utilizing the recently published fluorometric procedure of Faeder and Siegel (Faeder, E. J., and Siegle, L. M. (1973), Anal. Biochem. 53, 332) for the determination of FAD and FMN in mixtures, confirm the conclusions of Iyanagi and Mason for both rat and pig liver reductase preparations. Data for other flavoproteins are also presented. Iyanagi and Mason have also concluded that the air-stable "semiquinone" is a form of NADPH-cytochrome c reductase reduced by one electron per two falvines (F-FH). The present studies, however, do not agree with this conclusion, but instead support our previous results which indicate that both the aerobic and anaerobic half-reduced states of this flavoprotein exist in the two-electron reduced form (FH-FH). Removal of NADP+ does not affect the spectrum of the air-stable half-reduced form of the flavoprotein, nor does it affect the back titration of this intermediate by potassium ferricyanide. The possible implications of these observations on the catalytic cycle of the flavines of NADPH-cytochrome c reductase are discussed.     

The labial gland system of larvae of the imported fire ant,Solenopsis invicta Buren

Summary The authors describe the ultrastructure of the labial gland system in imported fire ant larvae, Solenopsis invicta Buren and present an enzyme analysis of enzymes in labial-gland secretions and midgut contents. The tubes of the labial gland produce and secrete a proteinaceous substance rich in digestive enzymes. The narrow cells of the reservoir of the gland have little or no secretory function but the lumen stores the proteinaceous secretion. The labial-gland enzymes include proteases and amylases, which function in extraintestinal digestion of solid food placed on the anteroventral body region of 4th-instar larvae by adult workers. The midgut contains proteases, amylases, and upases. Lipids appear to be predigested by the workers before being fed to the larvae.Approved as TA 15262 by the Director of the Texas Agricultural Experiment Station in cooperation with ARS/USDA. Supported by the Texas Department of Agriculure interagency agreement IAC-0487 (78-79)We would like to thank Rosemary Kamas, Dr. John Mirenda, and Mike Strand for their help in dissections of larvae and Dr. Howard Williams for his advice and suggestions     

Aerobic denitrification: a controversy revived

During studies on the denitrifying mixotroph, Thiosphaera pantotropha, it has been found that this organism is capable of simultaneously utilizing nitrate and oxygen as terminal electron acceptors in respiration. This phenomenon, termed aerobic denitrification, has been found in cultures maintained at dissolved oxygen concentrations up to 90% of air saturation.The evidence for aerobic denitrification was obtained from a number of independant experiments. Denitrifying enzymes were present even in organisms growing aerobically without nitrate. Aerobic yields on acetate were higher (8.1 g protein/mol) without than with (6.0 g protein/mol) nitrate, while the anaerobic yield with nitrate was even lower (4 g protein/mol). The maximum specific growth rate of Tsa. pantotropha was higher (0.34 h^-1) in the presence of both oxygen (>80% air saturation) and nitrate than in similar cultures not supplied with nitrate (0.27 h^-1), indicating that the rate of electron transport to oxygen was limiting. This was confirmed by oxygen uptake experiments which showed that although the rate of respiration on acetate was not affected by nitrate, the total oxygen uptake was reduced in its presence. The original oxygen uptake could be restored by the addition of denitrification inhibitors.Dedicated to Professor Dr. H.-G. Schlegel on the occasion of his 60th birthday     

Albert Howard and the mycorrhizal association

Albert Howard worked as an imperial agronomist for the British Government in India. Following his retirement in 1931, he returned to England and embarked on a passionate global campaign to reform agricultural practices. Central to Howard's project was the mycorrhizal association, a symbiotic relationship between plant roots and subterranean fungi, believed to play an important part in plant nutrition. I show that there are a number of close parallels between Howard's work in India and his portrayal of the mycorrhizal association, and argue that Howard used these fungi to naturalise his imperial project. Understood in this way, these mycorrhizal and imperial associations reveal ways that Howard was able to negotiate the boundaries between the local and global, England and India, science and agriculture, institute and village, and soil and plant. In contrast to Thomas Gieryn's work on hybridisation at the cultural boundaries between science and non-science, I concentrate on Howard's use of intermediaries to negotiate and articulate specific boundaries within his imperial project. Arguing that this approach reveals limitations in Gieryn's hybrid framework, I suggest that a focus on Howard's dependence on intermediaries draws attention to the discontinuities between entities, besides the dynamic ways that they might be coupled.     

Responses of lactic acid bacteria to oxygen

Abstract A small number of flavoprotein oxidase enzymes are responsible for the direct interaction of lactic acid bacteria (LAB) with oxygen; hydrogen peroxide or water are produced in these reactions. In some cultures exposed to oxygen, hydrogen peroxide accumulates to inhibitory levels.
Through these oxidase enzymes and NADH peroxidase, O₂ and H₂O₂ can accept electrons from sugar metabolism, and thus have a sparing effect on the use of metabolic intermediates, such as pyruvate or acetaldehyde, as electron acceptors. Consequently, sugar metabolism in aerated cultures of LAB can be substantially different from that in unaerated cultures. Energy and biomass yields, end-products of sugar metabolism and the range of substrates which can be metabolised are affected.
Lactic acid bacteria exhibit an inducible oxidative stress response when exposed to sublethal levels of H₂O₂. This response protects them if they are subsequently exposed to lethal concentrations of H₂O₂. The effect appears to be related to other stress responses such as heat-shock and is similar, in some but not all respects, to that previously reported for enteric bacteria.     

W. Montague Cobb (1904–1990): Physical Anthropologist, Anatomist, and Activist

William Montague Cobb's life and work reflect a profound integration of art, literature, social activism, and science. This article presents some of the highlights of his academic development and professional contributions. We have considered his early academic development within the contexts of the formative years of American physical anthropology, Howard University Medical School, and the social issues in American society that influenced Cobb. His approaches to teaching, anatomical and anthropological research, and medicine are unique, and yet are closely reasoned and creative reflections of the major currents of academe and the broader society with which he dealt. Imbued with a sense of social responsibility, Cobb's applied anthropology involved the accumulation of extensive data on the one hand, and the formation of organizations for social activism on the other. It was directed toward solving problems of health care and racism. His work thereby served to balance the widespread distortion and neglect of medical and racial problems facing A fro-America between 1930 and the present day. He was also a principal builder of black medical and scientific institutions, and he preserved the record of his coworkers' contributions through his many biographies. This work represents no more than a sketch of his rich and prolific career (during which he produced more than 1,100 publications); the emphasis of this biographical study has been to ascertain the circumstances and attitudes that helped mold the first Afro-American Ph.D. in physical anthropology.     

Gibberellin biosynthesis and the regulation of plant development

Gibberellins (GAs) form a large family of plant growth substances with distinct functions during the whole life cycle of higher plants. The rate of GA biosynthesis and catabolism determines how the GA hormone pool occurs in plants in a tissue and developmentally regulated manner. With the availability of genes coding for GA biosynthetic enzymes, our understanding has improved dramatically of how GA plant hormones regulate and integrate a wide range of growth and developmental processes. This review focuses on two plant systems, pumpkin and Arabidopsis, which have added significantly to our understanding of GA biosynthesis and its regulation. In addition, we present models for regulation of GA biosynthesis in transgenic plants, and discuss their suitability for altering plant growth and development.