Mass spectrometric based analysis of plasma membrane proteins that regulate cell expansion in Arabidopsis thaliana.

基于质谱分析调节拟南芥细胞扩张的质膜蛋白。

• 批准号: 1713899
• 负责人: Michael Sussman
• 金额: $ 45万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1713899&HistoricalAwards=false
• 关键词: Mass; spectrometric; based; analysis; plasma

Molecules present in the outer membranes of cells act as antennae to perceive changes in the environment, and transmit this information into the cell by chemical reactions (for example phosphorylation) and by changes in shape (or conformation.) Newly developed technologies allow such conformational changes in molecules to be detected using the methods of mass spectrometry, and in this project these new technologies will be used to understand how signals regulate the rapid expansion of plant cell size. Exercises will be developed for high school students, using mass spectrometry to analyze common food proteins such as soybean tofu. A PBS television series "Wednesday Nite at the Lab" that is broadcast free from the University of Wisconsin Biotechnology Center will explain to the general public (and demystify) mass spectrometry, using as an example the analysis of plant proteins and explaining how this impacts their everyday lives. Overall this project seeks to combine in vitro and in vivo approaches to understanding the molecular basis for signal transmission across the plasma membrane. The focus of this project is Feronia, a plant plasma membrane receptor kinase, and its peptide hormone ligand, RALF. When RALF binds to the extracellular domain of Feronia, the rate of cell expansion is reduced within minutes. This project will develop and apply novel mass spectrometric based tools for analyzing the conformational changes that extracellular RALF induces in Feronia. The newly devised PLIMB instrument (Plasma Induced Modification of Biomolecules) that generates microsecond pulses of hydroxyl radicals will be used to "footprint" Feronia and other proteins that are involved in Feronia/RALF action at the plasma membrane.

细胞外膜中存在的分子充当天线来感知环境的变化，并通过化学反应（例如磷酸化）和形状（或构象）的变化将这些信息传输到细胞中。新开发的技术允许使用质谱方法检测分子的构象变化，在这个项目中，这些新技术将用于了解信号如何调节细胞的快速扩张。 植物细胞大小。将为高中生开发练习，使用质谱分析大豆豆腐等常见食物蛋白质。威斯康星大学生物技术中心免费播出的 PBS 电视连续剧“实验室星期三之夜”将向公众解释（并揭开）质谱法，以植物蛋白分析为例，并解释这如何影响他们的日常生活。总体而言，该项目旨在结合体外和体内方法来了解跨质膜信号传输的分子基础。该项目的重点是 Feronia，一种植物质膜受体激酶及其肽激素配体 RALF。当 RALF 与 Feronia 的细胞外结构域结合时，细胞扩张速度在几分钟内就会降低。该项目将开发和应用基于质谱的新型工具来分析细胞外 RALF 在 Feronia 中诱导的构象变化。新设计的 PLIMB 仪器（等离子体诱导生物分子修饰）可产生微秒级羟基自由基脉冲，将用于“足迹”Feronia 和其他参与质膜上 Feronia/RALF 作用的蛋白质。

项目成果

A cell-free method for expressing and reconstituting membrane proteins enables functional characterization of the plant receptor-like protein kinase FERONIA

• DOI: 10.1074/jbc.m116.761981
• 发表时间: 2017-04-07
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Minkoff, Benjamin B.;Makino, Shin-ichi;Sussman, Michael R.
• 通讯作者: Sussman, Michael R.

Proteome-wide Analysis of Protein Thermal Stability in the Model Higher Plant Arabidopsis thaliana

模型高等植物拟南芥蛋白质热稳定性的全蛋白质组分析

• DOI: 10.1074/mcp.ra118.001124
• 发表时间: 2019
• 期刊: Molecular & Cellular Proteomics
• 影响因子: 7
• 作者: Volkening, Jeremy D.;Stecker, Kelly E.;Sussman, Michael R.
• 通讯作者: Sussman, Michael R.

Plasma-Generated OH Radical Production for Analyzing Three-Dimensional Structure in Protein Therapeutics

等离子体产生的 OH 自由基产生用于分析蛋白质治疗中的三维结构

• DOI: 10.1038/s41598-017
• 发表时间: 2017
• 期刊: Open access scientific reports
• 作者: Minkoff BB, Blatz JM

Function and solution structure of the Arabidopsis thaliana RALF8 peptide

• DOI: 10.1002/pro.3628
• 发表时间: 2019-06-01
• 期刊: PROTEIN SCIENCE
• 影响因子: 8
• 作者: Frederick, Ronnie O.;Haruta, Miyoshi;Markley, John L.
• 通讯作者: Markley, John L.

Environmental and Genetic Factors Regulating Localization of the Plant Plasma Membrane H+-ATPase

• DOI: 10.1104/pp.17.01126
• 发表时间: 2018-01-01
• 期刊: PLANT PHYSIOLOGY
• 影响因子: 7.4
• 作者: Haruta, Miyoshi;Tan, Li Xuan;Sussman, Michael R.
• 通讯作者: Sussman, Michael R.