Assembly of Cyanobacterial Carboxysomes

蓝藻羧基体的组装

• 批准号: 8686611
• 负责人: Myat T Lin
• 金额: $ 5.51万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8686611
• 关键词: Anabolism; Area; Bacteria; Bicarbonate Ion; Bicarbonate Ions; Biochemical Reaction; Biological; Biotechnology; Boxing; C-terminal; Capsid; Carbon Dioxide; Cell Nucleus; Chloroplasts; Complex; Confocal Microscopy; Cyanobacterium; Drug Delivery Systems; Electron Microscopy; Elements; Engineering; Enzymes; Escherichia coli; Eukaryota; Future; Galactosidase; Genes; Genome; Glycine; Goals; Image; Individual; Kinetics; Knowledge; Label; Metabolic; Methods; Molecular; Monitor; Nuclear; Organelles; Organism; Oxygenases; Pathway interactions; Peptide Signal Sequences; Peptides; Plant Leaves; Plants; Plastids; Play; Property; Proteins; Reaction; Regulation; Reporting; Ribulose-Bisphosphate Carboxylase; Role; Serine; Shapes; Structure; Synechococcus; Techniques; Thick; Tissues; Tobacco; Transgenic Organisms; Viral; Work; base; carbonate dehydratase; design; expression vector; macromolecular assembly; metabolic engineering; novel; prevent; promoter; protein expression; public health relevance; research study; sample fixation; vector

DESCRIPTION (provided by applicant): Many bacteria have a sophisticated mechanism for metabolic regulation that involves microcompartments. These bacterial microcompartments are made up of unique protein shells similar in structure to a viral capsid and enclose two or more enzymes in a particular reaction pathway. Carboxysomes from cyanobacteria and chemoauxotrophs are the most well known bacterial microcompartments. Within the last few years, the crystal structures of several shell components have been solved and have led to remarkable progress in the knowledge of subunit assembly and function of individual components. So far, there has been no report of heterologous assemblies of bacterial microcompartments in eukaryotic organisms. This project will target multiple carboxysome proteins into chloroplasts. This study will employ transient expression of foreign proteins fused with fluorescent tags by agroinfiltration technique. Any molecular complex assembled from the expressed proteins will be monitored with confocal microscopy, and the structures formed will be characterized by electron microscopy. In addition, this work will generate stable transgenic lines, in which carboxysome proteins are expressed from either the nucleus or the chloroplast genome. By expressing all necessary carboxysome structural components, this project will attempt to assemble the carboxysome microcompartment shells in eukaryotes. In addition, this study will determine whether it is possible to target foreign proteins to the carboxysome shells. The proposed study is an important step for engineering novel microcompartments and will have potential future applications in areas such as drug delivery, metabolic engineering and biotechnology.

描述（由申请人提供）：许多细菌具有涉及微型室的代谢调节的复杂机制。这些细菌的微型室由与病毒式衣壳相似的独特蛋白质壳组成，并在特定的反应途径中封闭了两个或多个酶。来自蓝细菌和化学巨人的羧化体是最著名的细菌微型室。在过去的几年中，已经解决了几个壳体成分的晶体结构，并导致了对单个组件的亚基组装和功能的显着进步。到目前为止，还没有关于真核生物中细菌微剖间异源组件的报道。该项目将将多种羧化体蛋白靶向叶绿体。这项研究将采用通过农业泄露技术与荧光标签融合的异物的短暂表达。从表达蛋白组装的任何分子复合物将通过共聚焦显微镜进行监测，形成的结构将以电子显微镜为特征。此外，这项工作将产生稳定的转基因线，其中羧化体蛋白从细胞核或叶绿体基因组中表达。通过表达所有必要的羧基结构组件，该项目将试图在真核生物中组装羧基小组壳。此外，这项研究将确定是否可以将异物蛋白靶向羧基壳。拟议的研究是工程新型微型院子的重要步骤，并将在药物递送，代谢工程和生物技术等领域具有潜在的未来应用。

项目成果

A faster Rubisco with potential to increase photosynthesis in crops.

• DOI: 10.1038/nature13776
• 发表时间: 2014-09-25
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Lin, Myat T.;Occhialini, Alessandro;Andralojc, P. John;Parry, Martin A. J.;Hanson, Maureen R.
• 通讯作者: Hanson, Maureen R.