植物活性氧的产生及其作用和危害

活性氧(ROS)是一类由O2转化而来的自由基或具有高反应活性的离子或分子。植物消耗的O2约有1%在叶绿体、线粒体、过氧化物酶体等多种亚细胞单位中被转化成了ROS。ROS有益或有害取决于它在植物体内的浓度。低浓度的ROS作为第二信使能在植物细胞信号转导途径中介导多种应答反应,高浓度的ROS则引起生物大分子的氧化损伤甚至细胞死亡。植物体内ROS产生和清除之间的平衡十分重要,并由一套有效的酶促和非酶促抗氧化系统来监控。该文主要系统介绍了植物ROS的种类、产生部位、在信号转导中的作用及其对植物细胞造成的主要伤害等方面的研究进展,为利用基因工程手段来提高植物对环境胁迫的抗性提供信息和思路。     

H2O2-NOX系统: 一种植物体内重要的发育调控与胁迫响应机制

以H₂O₂为中心的活性氧(reactive oxygen species, ROS)的产生是动植物发育与响应外界生物与非生物胁迫的普遍 特征, 其在生理和分子2个水平上调控植物的发育和对外界胁迫的响应, 并与一系列信号转导过程相关联。作为关键的ROS产生酶, 质膜NADPH氧化酶(plasma membrane NADPH oxidase, PM-NOX)在植物应对各种生物和非生物胁迫中具有重要作用, 被广泛认为是胁迫条件下植物细胞ROS产生并积累的主要来源。该文简要综述了近年来人们在植物细胞ROS产生、清除、生理功能以及PM-NOX酶的结构特征与功能等方面的研究进展, 并认为H₂O₂-NOX系统是一种植物体内普遍存在的重要发育调控与胁迫响应机制。     

H_2O_2-NOX系统：一种植物体内重要的发育调控与胁迫响应机制

以H2O2为中心的活性氧（reactive oxygen species,ROS）的产生是动植物发育与响应外界生物与非生物胁迫的普遍特征,其在生理和分子2个水平上调控植物的发育和对外界胁迫的响应,并与一系列信号转导过程相关联。作为关键的ROS产生酶,质膜NADPH氧化酶（plasma membrane NADPH oxidase,PM-NOX）在植物应对各种生物和非生物胁迫中具有重要作用,被广泛认为是胁迫条件下植物细胞ROS产生并积累的主要来源。该文简要综述了近年来人们在植物细胞ROS产生、清除、生理功能以及PM-NOX酶的结构特征与功能等方面的研究进展,并认为H2O2-NOX系统是一种植物体内普遍存在的重要发育调控与胁迫响应机制。     

气孔运动中的活性氧信号

大量研究证明活性氧(ROS)在气孔运动中起信号分子的作用。保卫细胞中ROS的产生依赖于特定的酶,其中NADPH氧化酶组分RBOH已得到深入研究,并已证实其参与生物与非生物胁迫反应。植物激素包括脱落酸(ABA)、水杨酸(SA)、乙烯、生长素及细胞分裂素等,它们均通过ROS的介导来调控气孔运动。生物胁迫(如毒性细菌和真菌)也会调控气孔运动。ROS参与这些调控过程。保卫细胞中存在多层次对ROS产生及其作用的调节,抗氧化活性物质和ROS敏感蛋白(如蛋白激酶和磷酸酶)均可传递ROS信号并调节气孔运动。ROS对离子通道调节的证据也越来越多。保卫细胞由于可通过ROS整合复杂的信号途径,已成为研究植物ROS信号转导过程的良好模式系统。     

气孔运动中的活性氧信号

大量研究证明活性氧(ROS)在气孔运动中起信号分子的作用。保卫细胞中ROS的产生依赖于特定的酶, 其中NADPH氧化酶组分RBOH已得到深入研究, 并已证实其参与生物与非生物胁迫反应。植物激素包括脱落酸(ABA)、水杨酸(SA)、乙烯、生长素及细胞分裂素等, 它们均通过ROS的介导来调控气孔运动。生物胁迫(如毒性细菌和真菌)也会调控气孔运动。ROS参与这些调控过程。保卫细胞中存在多层次对ROS产生及其作用的调节, 抗氧化活性物质和ROS敏感蛋白(如蛋白激酶和磷酸酶)均可传递ROS信号并调节气孔运动。ROS对离子通道调节的证据也越来越多。保卫细胞由于可通过ROS整合复杂的信号途径, 已成为研究植物ROS信号转导过程的良好模式系统。     

极端微生物及其应用研究进展

极端微生物是指在高/低温、高/低pH、高盐、高压等极端环境条件下生存的微生物.特殊的生存条件导致其具有特殊的遗传背景和代谢途径,并可产生功能特殊的酶类和活性物质.随着系统生物学和合成生物学技术的发展,极端微生物作为一类特殊的微生物群体,在生物医疗、生物能源和生物材料等领域具有巨大的应用潜力.极端微生物相关研究也对生命起...     

活性氧在植物非生物胁迫响应中功能的研究进展

活性氧(ROS)是植物在响应非生物胁迫过程中不可或缺的组成部分。适量的ROS可通过参与信号转导途径调节植物响应多种胁迫,而过量的ROS致使植物处于氧化应激状态。植物中每个亚细胞室都含有一套独立的ROS产生和清除途径,各自的ROS稳态水平及氧化还原状态也在不断发生变化,表现出各自独特的ROS特征。本文综述了近年来有关ROS在植物非生物胁迫响应过程中功能的研究进展及其在介导快速系统信号转导中的作用,为深入研究ROS在植物非生物胁迫响应中的功能提供参考。     

活性氧:从毒性分子到信号分子--活性氧与细胞的增殖、分化和凋亡及其信号转导途径

活性氧(reactive oxygen species,ROS)是生物体有氧代谢产生的一类活性含氧化合物的总称,主要包括O2·-、H2O2、·OH等,机体细胞通过多种途径维持ROS产生与消解的动态平衡。近年的研究揭示ROS参与细胞正常的生理过程,与细胞的增殖、分化及凋亡密切相关。不同刺激诱导细胞产生的内源性ROS可作为第二信使,通过改变氧化还原状态调节增殖、分化和凋亡相关的信号转导通路中多种靶分子的活性,最终决定细胞的命运。     

动物肠道菌群结构与代谢化学成分的相互关系

机体肠道对营养素的消化吸收以及抗病保健能力部分依赖于寄居其中的菌群结构。消化道正常菌群种类多、数量大,能产生多种酶类（消化酶和非消化酶类）,协助机体对营养素的消化吸收。此过程中可产生许多不同的化学成分（有益和有害成分）。本文主要综述机体肠道菌群结构与其内容物化学成分的相互关系。     

非吞噬细胞NAD(P)H氧化酶生成的活性氧参与基因表达和信号转导

以前认为,NAD(P)H氧化酶仅存在于吞噬细胞,负责吞噬细胞呼吸爆发时产生活性氧(ROS)以杀灭微生物。现在发现正常非吞噬细胞也有NAD(P)H氧化酶,称之为类NAD(P)H氧化酶。该酶在生长因子和细胞因子的刺激下,介导非吞噬细胞产生胞内或胞外的ROS,通过此途径产生的ROS对细胞增殖、分化和血管形成和缺氧反应等生理过程至关重要。这些新的发现。有力地证明了ROS作为细胞“信号分子”和“基因表达开关”的积极作用,改变了过去只把ROS看作有害物的误解。     

Cytosolic phospholipase A(2), lipoxygenase metabolites, and reactive oxygen species

Reactive oxygen species (ROS) are generated in mammalian cells via both enzymatic and non-enzymatic mechanisms. Although certain ROS production pathways are required for the performance of specific physiological functions, excessive ROS generation is harmful, and has been implicated in the pathogenesis of a number of diseases. Among the ROS-producing enzymes, NADPH oxidase is widely distributed among mammalian cells, and is a crucial source of ROS for physiological and pathological processes. Reactive oxygen species are also generated by arachidonic acid (AA) metabolites, which are released from membrane phospholipids via the activity of cytosolic phospholipase A(2) (cPLA(2)). In this study, we describe recent studies concerning the generation of ROS by AA metabolites. In particular, we have focused on the manner in which AA metabolism via lipoxygenase (LOX) and LOX metabolites contributes to ROS generation. By elucidating the signaling mechanisms that link LOX and LOX metabolites to ROS, we hope to shed light on the variety of physiological and pathological mechanisms associated with LOX metabolism.     

Detection and manipulation of mitochondrial reactive oxygen species in mammalian cells

Reactive oxygen species (ROS) are formed upon incomplete reduction of molecular oxygen (O₂) as an inevitable consequence of mitochondrial metabolism. Because ROS can damage biomolecules, cells contain elaborate antioxidant defense systems to prevent oxidative stress. In addition to their damaging effect, ROS can also operate as intracellular signaling molecules. Given the fact that mitochondrial ROS appear to be only generated at specific sites and that particular ROS species display a unique chemistry and have specific molecular targets, mitochondria-derived ROS might constitute local regulatory signals. The latter would allow individual mitochondria to auto-regulate their metabolism, shape and motility, enabling them to respond autonomously to the metabolic requirements of the cell. In this review we first summarize how mitochondrial ROS can be generated and removed in the living cell. Then we discuss experimental strategies for (local) detection of ROS by combining chemical or proteinaceous reporter molecules with quantitative live cell microscopy. Finally, approaches involving targeted pro- and antioxidants are presented, which allow the local manipulation of ROS levels.     

Regulation of Nox and Duox enzymatic activity and expression

In recent years, it has become clear that reactive oxygen species (ROS, which include superoxide, hydrogen peroxide, and other metabolites) are produced in biological systems. Rather than being simply a by-product of aerobic metabolism, it is now recognized that specific enzymes--the Nox (NADPH oxidase) and Duox (Dual oxidase) enzymes--seem to have the sole function of generating ROS in a carefully regulated manner, and key roles in signal transduction, immune function, hormone biosynthesis, and other normal biological functions are being uncovered. The prototypical Nox is the respiratory burst oxidase or phagocyte oxidase, which generates large amounts of superoxide and other reactive species in the phagosomes of neutrophils and macrophages, playing a central role in innate immunity by killing microbes. This enzyme system has been extensively studied over the past two decades, and provides a basis for comparison with the more recently described Nox and Duox enzymes, which generate ROS in a variety of cells and tissues. This review first considers the structure and regulation of the respiratory burst oxidase, and then reviews recent studies relating to the regulation of the activity of the novel Nox/Duox enzymes. The regulation of Nox and Duox expression in tissues and by specific stimuli is also considered here. An accompanying review considers biological and pathological roles of the Nox family of enzymes.     

Chromate-reducing properties of soluble flavoproteins from Pseudomonas putida and Escherichia coli

Cr(VI) (chromate) is a toxic, soluble environmental contaminant. Bacteria can reduce chromate to the insoluble and less toxic Cr(III), and thus chromate bioremediation is of interest. Genetic and protein engineering of suitable enzymes can improve bacterial bioremediation. Many bacterial enzymes catalyze one-electron reduction of chromate, generating Cr(V), which redox cycles, generating excessive reactive oxygen species (ROS). Such enzymes are not appropriate for bioremediation, as they harm the bacteria and their primary end product is not Cr(III). In this work, the chromate reductase activities of two electrophoretically pure soluble bacterial flavoproteins--ChrR (from Pseudomonas putida) and YieF (from Escherichia coli)-were examined. Both are dimers and reduce chromate efficiently to Cr(III) (kcat/Km = approximately 2 x 10(4) M(-1) x s(-1)). The ChrR dimer generated a flavin semiquinone during chromate reduction and transferred >25% of the NADH electrons to ROS. However, the semiquinone was formed transiently and ROS diminished with time. Thus, ChrR probably generates Cr(V), but only transiently. Studies with mutants showed that ChrR protects against chromate toxicity; this is possibly because it preempts chromate reduction by the cellular one-electron reducers, thereby minimizing ROS generation. ChrR is thus a suitable enzyme for further studies. During chromate reduction by YieF, no flavin semiquinone was generated and only 25% of the NADH electrons were transferred to ROS. The YieF dimer may therefore be an obligatory four-electron chromate reducer which in one step transfers three electrons to chromate and one to molecular oxygen. As a mutant lacking this enzyme could not be obtained, the role of YieF in chromate protection could not be directly explored. The results nevertheless suggest that YieF may be an even more suitable candidate for further studies than ChrR.     

Biotechnological approach of improving plant salt tolerance using antioxidants as markers

Salt stress causes multifarious adverse effects in plants. Of them, production of reactive oxygen species (ROS) is a common phenomenon. These ROS are highly reactive because they can interact with a number of cellular molecules and metabolites thereby leading to a number of destructive processes causing cellular damage. Plants possess to a variable extent antioxidant metabolites, enzymes and non-enzymes, that have the ability to detoxify ROS. In the present review, the emphasis of discussion has been on understanding the role of different antioxidants in plants defense against oxidative stress caused by salt stress. The role of different antioxidants as potential selection criteria for improving plant salt tolerance has been critically discussed. With the advances in molecular biology and availability of advanced genetic tools considerable progress has been made in the past two decades in improving salt-induced oxidative stress tolerance in plants by developing transgenic lines with altered levels of antioxidants of different crops. The potential of this approach in counteracting stress-induced oxidative stress has been discussed at length in this review.     

The influence of reactive oxygen species on cell cycle progression in mammalian cells

Cell cycle regulation is performed by cyclins and cyclin dependent kinases (CDKs). Recently, it has become clear that reactive oxygen species (ROS) influence the presence and activity of these enzymes and thereby control cell cycle progression. In this review, we first describe the discovery of enzymes specialized in ROS production: the NADPH oxidase (NOX) complexes. This discovery led to the recognition of ROS as essential players in many cellular processes, including cell cycle progression. ROS influence cell cycle progression in a context-dependent manner via phosphorylation and ubiquitination of CDKs and cell cycle regulatory molecules. We show that ROS often regulate ubiquitination via intermediate phosphorylation and that phosphorylation is thus the major regulatory mechanism influenced by ROS. In addition, ROS have recently been shown to be able to activate growth factor receptors. We will illustrate the diverse roles of ROS as mediators in cell cycle regulation by incorporating phosphorylation, ubiquitination and receptor activation in a model of cell cycle regulation involving EGF-receptor activation. We conclude that ROS can no longer be ignored when studying cell cycle progression.     

Nox enzymes, ROS, and chronic disease: an example of antagonistic pleiotropy

Reactive oxygen species (ROS) are considered to be chemically reactive with and damaging to biomolecules including DNA, protein, and lipid, and excessive exposure to ROS induces oxidative stress and causes genetic mutations. However, the recently described family of Nox and Duox enzymes generates ROS in a variety of tissues as part of normal physiological functions, which include innate immunity, signal transduction, and biochemical reactions, e.g., to produce thyroid hormone. Nature's "choice" of ROS to carry out these biological functions seems odd indeed, given its predisposition to cause molecular damage. This review describes normal biological roles of Nox enzymes as well as pathological conditions that are associated with ROS production by Nox enzymes. By far the most common conditions associated with Nox-derived ROS are chronic diseases that tend to appear late in life, including atherosclerosis, hypertension, diabetic nephropathy, lung fibrosis, cancer, Alzheimer's disease, and others. In almost all cases, with the exception of a few rare inherited conditions (e.g., related to innate immunity, gravity perception, and hypothyroidism), diseases are associated with overproduction of ROS by Nox enzymes; this results in oxidative stress that damages tissues over time. I propose that these pathological roles of Nox enzymes can be understood in terms of antagonistic pleiotropy: genes that confer a reproductive advantage early in life can have harmful effects late in life. Such genes are retained during evolution despite their harmful effects, because the force of natural selection declines with advanced age. This review discusses some of the proposed physiologic roles of Nox enzymes, and emphasizes the role of Nox enzymes in disease and the likely beneficial effects of drugs that target Nox enzymes, particularly in chronic diseases associated with an aging population.     

The molecular mechanism of protecting cells against oxidative stress by 2-selenium-bridged beta-cyclodextrin with glutathione peroxidase activity

Ultraviolet B (UVB) induces apoptosis and lipid peroxidation of NIH3T3 cells by producing reactive oxygen species (ROS). Glutathione peroxidase (GPX) is one of the most important antioxidant enzymes in organism and it can scavenge ROS. 2-selenium-bridged beta-cyclodextrin (2-SeCD) is a GPX mimic generated in our lab. Its GPX activity is 7.4 U/mumol, which is 7.5 times as much as that of ebselen. In this paper, we have established a cell damage system using UVB radiation. Using this system, we have determined antioxidant effect of 2-SeCD by comparison of malondialdehyde (MDA) and H(2)O(2) contents in NIH3T3 cells before and after UVB radiation. Experimental results indicate that 2-SeCD can inhibit lipid peroxidation and protect the cells from the damage generated by UVB radiation. To evaluate the molecular mechanism of this protection, we determined the effect of 2-SeCD on the expression of p53 and Bcl-2 in NIH3T3 cells. The results showed that 2-SeCD inhibits the increase of p53 expression level and the decrease of expression of Bcl-2 induced by UVB radiation. Thus, we have concluded that protection of NIH3T3 cells against oxidative stress by 2-SeCD was carried out by regulation of the expression of Bcl-2 and p53.     

Early steps in the DNA base excision/single-strand interruption repair pathway in mammalian cells

Base excision repair （BER） is an evolutionarily conserved process for maintaining genomic integrity by eliminating several dozen damaged （oxidized or aikylated） or inappropriate bases that are generated endogenously or induced by genotoxicants, predominantly, reactive oxygen species （ROS）. BER involves 4-5 steps starting with base excision by a DNA glycosylase, followed by a common pathway usually involving an AP-endonuclease （APE） to generate 3＇ OH terminus at the damage site, followed by repair synthesis with a DNA polymerase and nick sealing by a DNA iigase. This pathway is also responsible for repairing DNA single-strand breaks with blocked termini directly generated by ROS. Nearly all glycosylases, far fewer than their substrate lesions particularly for oxidized bases, have broad and overlapping substrate range, and could serve as back-up enzymes in vivo. In contrast, mammalian cells encode only one APE, APEI, unlike two APEs in lower organisms. In spite of overall similarity, BER with distinct subpathways in the mammals is more complex than in E. coli. The glycosylases form complexes with downstream proteins to carry out efficient repair via distinct subpathways one of which, responsible for repair of strand breaks with 3＇ phosphate termini generated by the NEIL family glycosylases or by ROS, requires the phosphatase activity of polynucleotide kinase instead of APE1. Different complexes may utilize distinct DNA polymerases and iigases. Mammalian glycosylases have nonconserved extensions at one of the termini, dispensable for enzymatic activity but needed for interaction with other BER and non-BER proteins for complex formation and organeile targeting. The mammalian enzymes are sometimes covalently modified which may affect activity and complex formation. The focus of this review is on the early steps in mammalian BER for oxidized damage.