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.

蛋白水解是蛋白质或肽对氨基酸的分解，对于去除不需要或受损的蛋白质以及细胞过程（例如代谢）至关重要。叶绿体是重要的植物细胞器，在生物量生产力和农业中很重要。然而，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种蛋白水解系统。这项研究将洞悉中央叶绿体CLP蛋白酶系统的作用，该系统由蛋白质伴侣，蛋白酶核心复合物以及参与特定CLP蛋白酶底物的选择和递送的几种蛋白质组成。研究结果也可以在分子种植和合成生物学（例如通过可调蛋白降解）中实施，因为诸如叶绿体等细胞室有利于过表达营养或药物价值的产品。这项研究的结果将允许更多理性蛋白质设计，以稳定地积累叶绿体蛋白，从而直接影响这些应用。该项目将提供有关蛋白质组学，质谱，分子遗传学，生物化学，本科，研究生和博士后水平的结构生物学的培训。我们的NSF赞助的REU计划将提供暑期实习。该项目建立在对生物材料（例如拟南芥蛋白酶突变体）的广泛投资上，以前被鉴定出的候选CLP底物或CLP适配器，以及质谱，蛋白质组学和结构生物学方面的专业知识。该建议的核心目的是揭示新型CLPS1-CLPF适配器系统的功能，结构和机械细节，包括底物选择，底物半衰期以及与CLP伴侣的相互作用。这也可能导致发现其他CLP适配器。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的高分辨率结构信息将用于CLPS1和CLPF及其功能伙伴。体外和植物相互作用分析和功能互补将决定蛋白质领域的作用。该奖项反映了NSF的法定任务，并且使用基金会的知识分子优点和更广泛的影响审查标准，被认为值得通过评估来获得支持。

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