Photosystem II as a model protein for understanding metalloenzyme active site assembly

光系统 II 作为理解金属酶活性位点组装的模型蛋白

• 批准号: 10300881
• 负责人: Christopher Gisriel
• 金额: $ 9.58万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10300881
• 关键词: Academia; Active Sites; Address; Anabolism; Binding; Binding Sites; Biochemical; Biogenesis; Biological Models; Biology; Cations; Cells; Characteristics; Chemistry; Complex; Computer Models; Coupled; Dependence; Development; Engineering; Environment; Enzymes; Event; Exhibits; Goals; In Vitro; Investigation; Ions; Knowledge; Lead; Light; Location; Maintenance; Metal Ion Binding; Metals; Methods; Modeling; Molecular; Molecular Conformation; Molecular Structure; Nature; Oxidation-Reduction; Oxides; Oxidoreductase; Oxygen; Pathway interactions; Phase; Positioning Attribute; Preparation; Process; Protein Complex Subunit; Protein Conformation; Proteins; Recombinant Proteins; Research; Research Personnel; Resolution; Role; Science; Series; Structure; Structure-Activity Relationship; Techniques; Therapeutic; Training; Water; Work; base; carboxylate; career; cofactor; design; human disease; improved; in vivo; insight; metalloenzyme; novel therapeutics; oxidative damage; photoactivation; photosystem II; polypeptide; repaired

Project Summary Metalloenzymes are ubiquitous throughout biology and implicated in a variety of human diseases. A gap in our understanding of metalloenzymes lies in their maturation. Biogenesis intermediates precede formation of the mature enzyme and must be tightly regulated to correctly assemble the metallocofactor in the active site. Despite the wealth of knowledge on mature metalloenzymes, their biogenesis, especially active site assembly, is not well understood. Photosystem II (PSII) is a light-driven oxidoreductase whose active site contains a metallocofactor that is assembled through a series of step-wise ion binding and photooxidation events called photoactivation. Photoactivation provides a convenient model for understanding metallocofactor assembly because it is facile; in vitro assembly may be achieved by simply adding the required ions in solution and assembly is advanced by providing light quanta. Furthermore, recent advances in structural investigation of PSII have developed a platform for routinely solving high-resolution structures of PSII assembly intermediates. We propose to characterize biogenesis intermediates of PSII that may reveal generalized rules for metalloenzyme assembly. The long-term goals of this project are to provide structural and functional bases for assembly of the metallocofactor in PSII (training phase), and reveal how PSII biogenesis intermediates maintain precursor structures important for active site maintenance (independent phase). We will use structural approaches, biochemical techniques, and computational modeling to achieve the following three specific objectives: (1) to reveal how the oxygen evolving complex of PSII is assembled, (2) to uncover precursor structures of PSII that are integral for assembling its active site, and (3) to understand the role of transiently-bound subunits during PSII biogenesis. Gaining such insight may allow for the development of new therapeutics and design principles for de novo metalloenzyme engineering.

项目摘要 金属酶在生物学中普遍存在，并与多种人类疾病有关。我们之间的差距 对金属酶的理解在于它们的成熟。生物发生中间体在形成之前 成熟的酶，必须严格调节，以正确组装的金属辅因子的活性位点。尽管 关于成熟金属酶、它们的生物起源，特别是活性位点组装的丰富知识还不是很好 明白光系统II（PSII）是一种光驱动的氧化还原酶，其活性位点含有金属辅因子 其通过一系列逐步离子结合和称为光活化的光氧化事件组装。 光活化为理解金属辅因子组装提供了一个方便的模型，因为它是容易的; 体外组装可以通过简单地在溶液中加入所需的离子来实现 提供光量子。此外，PSII结构研究的最新进展已经发展了一种新的 平台，用于常规求解PSII组装中间体的高分辨率结构。我们建议 表征PSII的生物合成中间体，可能揭示金属酶组装的一般规则。 该项目的长期目标是为装配 金属辅因子在PSII（训练阶段），并揭示如何PSII生物合成中间维持前体 对现场维护很重要的结构（独立阶段）。我们将使用结构化方法， 生物化学技术和计算建模，以实现以下三个具体目标：（1） 揭示PSII的氧释放复合物是如何组装的，（2）揭示PSII的前体结构， 是组装其活性位点不可或缺的，（3）了解瞬时结合亚基在PSII过程中的作用 生物起源。获得这样的洞察力可以允许开发新的治疗方法和设计原则， 从头金属酶工程。