Novel Responses to Oxygen by Anaerobic Microorganisms
厌氧微生物对氧气的新反应
基本信息
- 批准号:7025621
- 负责人:
- 金额:$ 26.95万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2000
- 资助国家:美国
- 起止时间:2000-03-01 至 2009-02-28
- 项目状态:已结题
- 来源:
- 关键词:NAD(P)H dehydrogenaseRaman spectrometryX ray crystallographyactive sitesaerobiosisanaerobic bacteriaanaerobiosiscatalystcircular magnetic dichroismelectron nuclear double resonance spectroscopyelectron spin resonance spectroscopyenzyme activityenzyme complexenzyme mechanismfree radical oxygenfunctional /structural genomicshydrogen peroxideinfrared spectrometryiron sulfur proteinmicroarray technologyoxidative stressoxidoreductaseprotein structure functionrespiratory burst oxidasestress proteinssuperoxides
项目摘要
DESCRIPTION (provided by applicant): By definition, aerobic organisms, both prokaryotic and eukaryotic, require molecular oxygen for energy conservation. This is a mixed blessing, however, as extremely reactive oxygen derivatives are produced during normal metabolism that can damage all cellular components. These so-called reactive oxygen species (ROS) have been implicated in a wide variety of chronic and infectious human diseases, including cancer, Alzheimer's disease, arthritis and AIDS, yet ROS are also used as a defense system against pathogens and in signal transduction pathways. Understanding the responses of microbes to oxygen has direct ramifications for the treatment of diseases caused by anaerobic pathogens. In 1999 we proposed that anaerobes have a novel response to ROS in which a non-heme iron protein termed superoxide reductase (SOR) played a key role. SOR was characterized from the hyperthermophilic anaerobe, Pyrococcus furiosus, and over the prior funding period it has been established using structural and spectroscopic approaches that SOR is uniquely suited to catalyze superoxide reduction. Using DNA microarrays to all 2065 ORFs in the complete P. furiosus genome, it was shown that the genes encoding SOR and related proteins are all expressed at significant levels in the absence of any oxidative shock. P. furiosus is therefore continuously 'armed' and ready to deal with ROS exposure. This is a first line of defense, however, as DNA microarray analyses show that the complete response to oxidative stress requires the induction of a large number of novel proteins (encoded by conserved/hypothetical genes), some of which are also induced by growth at sub-optimal temperatures. In the proposed research, the novel stress-regulated proteins, together with SOR and related reductases and oxidases, will be characterized with respect to their regulation, multiprotein complex formation, and catalytic functions using immunological, biochemical and structural analyses. A variety of complementary spectroscopic techniques, including EPR, ENDOR, MCD, resonance Raman, FTIR and X-ray absorption, will be utilized to probe the catalytic function of specific members of the stress-related pathways, with particular emphasis on SOR. The results will provide completely new insights into the stress responses of anaerobes, and provide strategies for determining the function of uncharacterized hypothetic algenes that typically account for half of a microbial genome.
描述(由申请人提供):根据定义,原核和真核的有氧生物需要分子氧来节能。但是,这是一种混合的祝福,因为在正常代谢过程中产生了极其活性的氧衍生物,从而损害了所有细胞成分。这些所谓的活性氧(ROS)已与多种慢性和感染性人类疾病有关,包括癌症,阿尔茨海默氏病,关节炎和艾滋病,但ROS也被用作对病原体和信号转导途径的防御系统。了解微生物对氧气的反应具有直接的影响,以治疗由厌氧病原体引起的疾病。在1999年,我们提出,厌氧菌对ROS有一种新颖的反应,其中非血红素蛋白称为超氧化物还原酶(SOR)起着关键作用。 SOR的特征是通过使用结构和光谱方法建立的,在先前的资金期间,在先前的资金期间,SOR的特征是SOR非常适合催化超氧化物的还原。使用DNA微阵列到完整的狂热假单胞菌基因组中的所有2065个ORF,结果表明,在没有任何氧化休克的情况下,编码SOR和相关蛋白的基因都在显着水平上表达。因此,Furiosus持续“武装”,并准备应对ROS暴露。然而,这是第一道防线,因为DNA微阵列分析表明,对氧化应激的完全反应需要诱导大量新型蛋白质(由保守/假设基因编码),其中一些也是由亚次优等温度下的生长引起的。在拟议的研究中,新型应激调节的蛋白质以及SOR和相关还原酶和氧化酶的调节,多蛋白复合物的形成以及使用免疫学,生化和结构分析的催化功能的表征。将利用多种互补的光谱技术,包括EPR,Endor,MCD,Resonance Raman,FTIR和X射线吸收,用于探测与压力相关途径的特定成员的催化功能,并特别强调SOR。结果将为厌氧菌的应力反应提供全新的见解,并提供了确定通常占微生物基因组一半的未表征的假设Algenes功能的策略。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Michael W. Adams其他文献
Michael W. Adams的其他文献
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