CLPS1-CLPF ADAPTORS FOR PROTEASE SUBSTRATE SELECTION IN CHLOROPLASTS

用于叶绿体中蛋白酶底物选择的 CLPS1-CLPF 接头

基本信息

批准号：
1940961
负责人：
Klaas van Wijk
金额：
$ 93万
依托单位：
Cornell University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1940961&HistoricalAwards=false
关键词：
CLPS1 CLPF ADAPTORS PROTEASE SUBSTRATE

项目摘要

Proteolysis, the breakdown of proteins or peptides to amino acids, is critical for removal of unwanted or damaged proteins and regulation of cellular processes such as metabolism. Chloroplasts are essential plant organelles that are important in biomass productivity and agriculture. However, the determinants of chloroplast protein life-time and degradation are poorly understood. Cells and organelles contain hundreds of proteolytic systems. This research will provide insight into the role of the central chloroplast Clp protease system, consisting of protein chaperones, a protease core complex and several proteins involved in selection and delivery of specific Clp protease substrates. The research findings can also be implemented in molecular farming and synthetic biology (e.g. through tunable protein degradation), since cellular compartments such as chloroplasts are favored for overexpression of products with nutritional or pharmaceutical value. The outcome of this research will allow more rational protein design for stable accumulation of chloroplast proteins, thus directly impacting these applications. This project will provide training in proteomics, mass spectrometry, molecular genetics, biochemistry, structural biology at the undergraduate, graduate and post-doctoral levels. Summer internships will be offered through our NSF-sponsored REU programs. This project is built on extensive investments in biological materials such as Arabidopsis protease mutants, previously identified candidate Clp substrate or Clp adaptors, as well as expertise in mass spectrometry, proteomics and structural biology. The central objective of this proposal is to unravel the functional, structural, and mechanistic details of a novel ClpS1-ClpF adaptor system, including substrate selection, substrate half-life, and interactions with Clp chaperones. It may also lead to the discovery of additional Clp adaptors. We will use our in vivo tools to test the physiological role of ClpS1 and ClpF in substrate selection. In vivo degradation assays will test the impact of Clp components on the half-life of candidate substrates. High-resolution structural information through X-ray crystallography or CryoEM will be obtained for ClpS1 and ClpF, with their functional partners. In vitro and in planta interaction assays and functional complementation will determine the role of protein domains.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

蛋白水解，即蛋白质或肽分解为氨基酸，对于去除不需要或受损的蛋白质以及调节新陈代谢等细胞过程至关重要。叶绿体是重要的植物细胞器，对生物质生产力和农业非常重要。然而，人们对叶绿体蛋白寿命和降解的决定因素知之甚少。细胞和细胞器包含数百个蛋白水解系统。这项研究将深入了解中央叶绿体 Clp 蛋白酶系统的作用，该系统由蛋白质伴侣、蛋白酶核心复合物和参与特定 Clp 蛋白酶底物选择和传递的几种蛋白质组成。研究结果还可以应用于分子农业和合成生物学（例如通过可调节的蛋白质降解），因为叶绿体等细胞区室有利于过度表达具有营养或药物价值的产品。这项研究的结果将允许更合理的蛋白质设计，以实现叶绿体蛋白质的稳定积累，从而直接影响这些应用。该项目将为本科生、研究生和博士后提供蛋白质组学、质谱、分子遗传学、生物化学、结构生物学方面的培训。暑期实习将通过我们的 NSF 赞助的 REU 项目提供。该项目建立在对生物材料（如拟南芥蛋白酶突变体、先前确定的候选 Clp 底物或 Clp 接头）以及质谱、蛋白质组学和结构生物学方面的专业知识的广泛投资的基础上。该提案的中心目标是阐明新型 ClpS1-ClpF 接头系统的功能、结构和机制细节，包括底物选择、底物半衰期以及与 Clp 伴侣的相互作用。它还可能导致其他 Clp 适配器的发现。我们将使用我们的体内工具来测试 ClpS1 和 ClpF 在底物选择中的生理作用。体内降解测定将测试 Clp 成分对候选底物半衰期的影响。通过 X 射线晶体学或 CryoEM 可以获得 ClpS1 和 ClpF 及其功能伙伴的高分辨率结构信息。体外和植物内相互作用测定和功能互补将决定蛋白质结构域的作用。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The CLP and PREP protease systems coordinate maturation and degradation of the chloroplast proteome in Arabidopsis thaliana

CLP 和 PREP 蛋白酶系统协调拟南芥叶绿体蛋白质组的成熟和降解

DOI：
10.1111/nph.18426
发表时间：
2022
期刊：
New Phytologist
影响因子：
9.4
作者：
Rowland, Elden;Kim, Jitae;Friso, Giulia;Poliakov, Anton;Ponnala, Lalit;Sun, Qi;van Wijk, Klaas J.
通讯作者：
van Wijk, Klaas J.

Autocatalytic Processing and Substrate Specificity of Arabidopsis Chloroplast Glutamyl Peptidase