Assembly and Repair of Biological Iron-Sulfur Clusters

生物铁硫簇的组装和修复

• 批准号: 9315821
• 负责人: MICHAEL K. JOHNSON
• 金额: $ 32.4万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9315821
• 关键词: Active Sites; Address; Aerobic; Aging-Related Process; Anabolism; Anemia; Archaea; Ataxia; Atherosclerosis; Bacteria; Biochemical; Biogenesis; Biological; Biological Assay; Biophysics; Carbon; Carrier Proteins; Chloroplasts; Circular Dichroism; Complement; Complex; Coupled; Cyanobacterium; Defect; Disease; Electron Spin Resonance Spectroscopy; Electron Transport; Enzymes; Eukaryota; Fumarates; Goals; Health; Homeostasis; Human; Hydrogen; In Situ; In Vitro; Iron; Iron Overload; Iron-Sulfur Proteins; Life; Ligation; Magnetism; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metabolism; Mitochondria; Molecular; Molecular Biology; Molecular Biology Techniques; Myopathy; Nature; Neurodegenerative Disorders; Nitrates; Nitrogen; Nitrogen Fixation; Organism; Oxidation-Reduction; Oxidative Stress; Oxygen; Plants; Plastids; Process; Property; Prosthesis; Proteins; Reactive Oxygen Species; Regulation; Respiratory Chain; Role; Scaffolding Protein; Signal Transduction; Site; Specificity; Sulfur; System; TXN gene; Techniques; Temperature; Work; absorption; age related; base; biophysical techniques; circular magnetic dichroism; cysteine desulfurase; design; experimental study; genetic regulatory protein; glutaredoxin; human disease; in vivo; iron deficiency; oxidative damage; pathogenic bacteria; public health relevance; repaired; respiratory; stem; trafficking

DESCRIPTION (provided by applicant): Iron-sulfur clusters are present in more than 300 different types of enzymes or proteins and constitute one of the most ancient, ubiquitous and structurally diverse classes of biological prosthetic groups. However, the most common type of cluster, the cubane-type [Fe4S4] cluster, is particularly sensitive to oxidative degradation. Hence, the process of iron-sulfur biosynthesis and repair is essential to almost all aerobic life forms and is remarkably conserved in prokaryotic and eukaryotic organisms. Three distinct types of iron-sulfur cluster assembly machinery have emerged in bacteria, termed the NIF, ISC and SUF systems, and the ISC and SUF systems form the basis of the eukaryotic mitochondrial and plastid iron-sulfur cluster assembly machineries, respectively. In each case the overall mechanism involves cysteine desulfurase-mediated assembly of transient clusters on scaffold proteins and subsequent transfer of preformed clusters or cluster fragments to apo proteins. However, in no case is the assembly, repair or transfer mechanism understood at the molecular level. The long-term goal of this project is a molecular-level understanding of iron-sulfur cluster biosynthesis and repair using the NIF, ISC and SUF systems, and accessory proteins such as monothiol glutaredoxins and thioredoxin-like Nfu proteins. Elucidating the mechanism of iron-sulfur cluster biosynthesis and repair is central to understanding cellular iron homeostasis and thereby human diseases associated with iron-overload, oxidative stress and defects in the mitochondrial respiratory chain. The approach involves using molecular biology techniques to effect large scale expression and/or site-specific changes in the target enzymes and proteins, biochemical and enzymatic assays, and the application of biophysical spectroscopic techniques (electron paramagnetic resonance, UV-visible absorption, circular dichroism, and magnetically-induced circular dichroism, resonance Raman, M�ssbauer, and mass spectrometry) that can probe the nature, ligation and detailed properties of iron or iron-sulfur centers during cluster biosynthesis, degradation, repair or transfer to acceptor proteins. The objectives are to establish the molecular mechanisms of assembly, degradation, repair, and transfer of iron-sulfur clusters, the specificity of cluster transfer with respect to acceptor proteins, and the means by which iron-sulfur proteins regulate cellular iron homeostasis.

描述（通过应用程序）：硫磺簇在300多种类型的酶，无处不在，结构上多样化的生物假体类别。生命形式和我的真核生物非常保守。在预先形成的簇或簇片段O apo蛋白的脚手架和随后的转移时，在分子水平上的组装，修复或转移机械中， 使用NIF，ISC和SUF系统的生物合成和修复，以及辅助蛋白，例如Monithins和Thioredoxin。在该方法中，涉及使用分子生物学技术来促进靶酶和蛋白质中的大规模表达和/或细节变化，生化和酶促测定生物物理学光谱技术ROROSIST ROROISM，RESONANCE RAMAN，M.SSSBABAUER和MOSS SBABAUER和质量特殊光谱仪）在簇生物合成，降解，修复或转移对受体蛋白的过程中，铁或铁硫中心的性质，结扎和详细特性是建立目标。 铁硫簇PTOR蛋白的组装，修复和转移的分子机制，以及铁硫蛋白调节细胞铁稳态的手段。