Assembly of Biological Iron-Sulfur Clusters

生物铁硫簇的组装

• 批准号: 8240101
• 负责人: MICHAEL K. JOHNSON
• 金额: $ 28.38万
• 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 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8240101
• 关键词: Active Sites; Address; Anabolism; Anemia; Ataxia; Attention; Azotobacter vinelandii; Binding; Biochemical; Biogenesis; Biological; Biological Assay; Carbon; Chloroplasts; Circular Dichroism; Complement; Coupled; Crystallography; Defect; Disease; Disulfides; Electron Spin Resonance Spectroscopy; Electron Transport; Enzymes; Generations; Goals; Health; Homeostasis; Human; Hydrogen; In Vitro; Iron; Iron Overload; Iron-Sulfur Proteins; Life; Magnetism; Mass Spectrum Analysis; Mediating; Mediation; Metabolism; Mitochondria; Molecular; Molecular Biology; Molecular Biology Techniques; Myopathy; Nature; Nitrogen; Nitrogen Fixation; Nitrogen Fixation Genes; Organism; Oxidation-Reduction; Oxidoreductase; Oxygen; Plant Proteins; Process; Property; Prosthesis; Protein S; Proteins; Regulation; Respiratory Chain; Role; S-Adenosylmethionine; Scaffolding Protein; Signal Transduction; Site; Specificity; Staging; Structure; Sulfides; Sulfur; System; Techniques; Temperature; Thioredoxin; Work; X-Ray Crystallography; absorption; base; circular magnetic dichroism; cysteine desulfurase; design; glutaredoxin; human disease; in vivo; iron deficiency; novel; polypeptide; repaired; research study; respiratory; stem

DESCRIPTION (provided by applicant): Project Summary Iron-sulfur clusters are present in more than 200 different types of enzymes or proteins and constitute one of the most ancient, ubiquitous and structurally diverse classes of biological prosthetic groups. Hence the process of iron-sulfur biosynthesis is essential to almost all forms of life and is remarkably conserved in prokaryotic and eukaryotic organisms. Three distinct types of iron-sulfur cluster assembly machinery have emerged, termed the NIF, ISC and SUF systems, and 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 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 using the NIF, ISC and SUF systems. Elucidating the mechanism of iron-sulfur cluster biosynthesis is central to understanding cellular iron homeostasis and thereby human diseases associated with iron-overload 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, X-ray crystallography, and the application of biophysical spectroscopic techniques (electron paramagnetic resonance, absorption, magnetic and natural circular dichroism, resonance Raman, and M"ssbauer) that can probe the nature and detailed properties of iron or iron-sulfur centers during cluster biosynthesis or transfer to acceptor proteins. The objectives are to establish the structure of cluster-bound forms and the molecular mechanism of cluster assembly and transfer for each the four known types of iron-sulfur cluster scaffold proteins, identify the roles and specificity of each type of scaffold protein in the maturation of iron-sulfur proteins, and characterize the mechanisms used to regulate iron-sulfur cluster biosynthesis. PUBLIC HEALTH RELEVANCE: The importance of iron-sulfur clusters to human health stems largely from their crucial role in iron homeostasis and their involvement in a large number of enzymes and proteins, particularly those in the mitochondrial respiratory chain. A molecular-level understanding of iron-sulfur cluster biogenesis is crucial for understanding a variety of human diseases involving anemias, myopathies and ataxias that arise from defects in iron-sulfur cluster assembly proteins.

描述（由申请人提供）： 项目摘要铁硫簇存在于200多种类型的酶或蛋白质中，构成了最古老，无处不在和结构上多样化的生物假体群体之一。因此，铁硫生物合成的过程对于几乎所有形式的生命都是必不可少的，在原核和真核生物中是明显的。已经出现了三种不同类型的铁硫簇组装机械机械，称为NIF，ISC和SUF系统，在每种情况下，总体机制都涉及半胱氨酸脱硫酶介导的瞬态簇在脚手架蛋白上的瞬态簇的组装，并随后转移的前提簇或群集片段的转移到Apo蛋白。但是，在任何情况下，在分子水平上都理解了组装或转移机制。该项目的长期目标是使用NIF，ISC和SUF系统对铁硫簇生物合成的分子水平理解。阐明铁硫簇生物合成的机制对于理解细胞铁稳态是至关重要的，从而在线粒体呼吸链中与铁超负荷和缺陷有关的人类疾病。该方法涉及使用分子生物学技术来实现靶酶和蛋白质，生化和酶测定，X射线晶体学的大规模表达和/或位点特异性变化，并应用生物物理光谱技术（电子磁磁共振，吸收，吸收，吸收，吸收，吸收，吸收，吸收）磁性和天然圆形二色性，共振拉曼和m“ ssbauer）可以在簇生物合成期间探测铁或铁硫中心的性质和详细特性或转移到受体蛋白上。这些目标是建立群集结合形式的结构以及每种已知类型的铁硫簇支架蛋白的簇组装和转移的分子机制，确定每种类型的支架蛋白在铁硫蛋白成熟中的作用和特异性，并表征用于调节的机制铁硫簇生物合成。对铁硫簇生物发生的分子水平理解对于理解涉及贫血，肌病和共济失调的各种人类疾病至关重要，这些疾病是由铁硫簇组装蛋白缺陷引起的。

项目成果

Identification of the Ferredoxin-Binding Site of a Ferredoxin-Dependent Cyanobacterial Nitrate Reductase.

• DOI: 10.1021/acs.biochem.7b00025
• 发表时间: 2017-10-17
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Srivastava AP;Hardy EP;Allen JP;Vaccaro BJ;Johnson MK;Knaff DB
• 通讯作者: Knaff DB

Iron-sulfur cluster biosynthesis.

• DOI: 10.1042/bst0361112
• 发表时间: 2008-12
• 期刊: Biochemical Society transactions
• 影响因子: 3.9
• 作者: Bandyopadhyay S;Chandramouli K;Johnson MK
• 通讯作者: Johnson MK

Glutaredoxins: roles in iron homeostasis.

• DOI: 10.1016/j.tibs.2009.08.005
• 发表时间: 2010-01
• 期刊: TRENDS IN BIOCHEMICAL SCIENCES
• 影响因子: 13.8
• 作者: Rouhier, Nicolas;Couturier, Jeremy;Johnson, Michael K.;Jacquot, Jean-Pierre
• 通讯作者: Jacquot, Jean-Pierre

Role of conserved cysteines in mediating sulfur transfer from IscS to IscU.

保守半胱氨酸在介导硫从 IscS 转移到 IscU 中的作用。

• DOI: 10.1016/j.febslet.2005.08.046
• 发表时间: 2005
• 期刊: FEBS letters.
• 作者: Smith,ArcherD;Frazzon,Jeverson;Dean,DennisR;Johnson,MichaelK
• 通讯作者: Johnson,MichaelK

Characterization of [4Fe-4S]-containing and cluster-free forms of Streptomyces WhiD.

• DOI: 10.1021/bi901498v
• 发表时间: 2009-12-29
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Crack, Jason C.;den Hengst, Chris D.;Jakimowicz, Piotr;Subramanian, Sowmya;Johnson, Michael K.;Buttner, Mark J.;Thomson, Andrew J.;Le Brun, Nick E.
• 通讯作者: Le Brun, Nick E.