Collaborative Research: Heme Distortion and Protein-Protein Contacts in Oxygen-Dependent Globin Coupled Sensor Signaling

合作研究：氧依赖性珠蛋白耦合传感器信号中的血红素畸变和蛋白质-蛋白质接触

• 批准号: 2003350
• 负责人: Emily Weinert
• 金额: $ 49.72万
• 依托单位: Pennsylvania State Univ University Park
• 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: 2020-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2003350&HistoricalAwards=false
• 关键词: Collaborative; Research; Heme; Distortion; Protein

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Emily Weinert from Pennsylvania State University, Dr. Christine Dunham from Emory University, and Dr. James Kincaid from Marquette University to investigate bacterial oxygen-sensing proteins. Bacteria are integral to the health of ecosystems, plants, animals, and humans, and have evolved to adapt to constantly changing conditions within their environment. However, the mechanisms by which bacteria sense and respond to changes in the levels of environmental gases are poorly understood. The research supported by this award examines the roles of environmental signals in controlling bacterial phenotypes; more specifically it examines the signaling mechanism and pathway of a family of proteins termed globin-coupled sensors that are responsible for sensing oxygen. This project provides specialized training for students in protein chemistry, spectroscopy, and X-ray crystallography. Additionally, the participants in this project are integrating their research results into a board game to create lesson plans that make elementary and middle school students aware of how proteins help bacteria sense and react to changes in their environment. This research project seeks to characterize the intra- and inter-molecular protein interactions that control oxygen-dependent signaling by a family of bacterial heme proteins termed globin-coupled sensors. Globin-coupled sensor proteins are widely distributed within bacteria and contain output domains that are linked to important phenotypes, including biofilm formation, motility, and virulence. A combination of research methods that includes mutagenesis/screening, enzyme kinetics, resonance Raman spectroscopy, and X-ray crystallography is being used to interrogate the oxygen-dependent signaling mechanism within globin coupled sensor proteins and the transmission of the signaling events to downstream partners. Results from this study are expected to provide new information about bacterial gas-sensing proteins, signal transduction pathways, and regulation of key bacterial enzymatic domains involved in controlling biofilm formation and virulence.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.

通过这一奖项，化学系的生命过程化学计划资助宾夕法尼亚州立大学的Emily Weinert博士、Emory大学的Christine Dunham博士和Marquette大学的James Kincaid博士研究细菌氧感应蛋白质。细菌对生态系统、植物、动物和人类的健康是不可或缺的，并已进化为适应环境中不断变化的条件。然而，细菌感知和响应环境气体水平变化的机制却知之甚少。这项由该奖项支持的研究检查了环境信号在控制细菌表型中的作用；更具体地说，它检查了一系列被称为珠蛋白偶联传感器的蛋白质的信号机制和途径，这些蛋白质负责感知氧气。该项目为学生提供蛋白质化学、光谱学和X射线结晶学方面的专门培训。此外，该项目的参与者正在将他们的研究成果整合到一个棋盘游戏中，以创建教案，让中小学生意识到蛋白质如何帮助细菌感知和反应环境的变化。这项研究项目试图通过一类被称为珠蛋白偶联传感器的细菌血红素蛋白家族来表征控制氧依赖信号的分子内和分子间蛋白质相互作用。珠蛋白偶联传感器蛋白在细菌中广泛分布，包含与重要表型有关的输出域，包括生物膜形成、运动性和毒力。一系列研究方法，包括突变/筛选、酶动力学、共振拉曼光谱和X射线结晶学，正被用来询问珠蛋白偶联传感器蛋白内的氧依赖信号机制以及信号事件向下游伙伴的传递。这项研究的结果有望提供有关细菌气敏蛋白、信号转导途径以及控制生物膜形成和毒力的关键细菌酶结构域调控的新信息。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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

Differential ligand-selective control of opposing enzymatic activities within a bifunctional c-di-GMP enzyme

• DOI: 10.1073/pnas.2100657118
• 发表时间: 2021-09
• 期刊: Proceedings of the National Academy of Sciences
• 作者: Dayna C. Patterson;Myrrh Perez Ruiz;H. Yoon;Johnnie A. Walker;J. Armache;N. Yennawar;Emily E. Weinert

Heme-Edge Residues Modulate Signal Transduction within a Bifunctional Homo-Dimeric Sensor Protein

血红素边缘残基调节双功能同源二聚体传感器蛋白内的信号转导

• DOI: 10.1021/acs.biochem.1c00581
• 发表时间: 2021
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Patterson, Dayna C.;Liu, Yilin;Das, Sayan;Yennawar, Neela H.;Armache, Jean-Paul;Kincaid, James R.;Weinert, Emily E.
• 通讯作者: Weinert, Emily E.

Spectrophotometric Method for the Quantification and Kinetic Evaluation of in vitro c-di-GMP Hydrolysis in the Presence and Absence of Oxygen

有氧和无氧情况下体外 c-di-GMP 水解的定量和动力学评价的分光光度法

• 发表时间: 2023
• 期刊: Methods in molecular biology
• 作者: Patterson, Dayna C.;Weinert, Emily E.
• 通讯作者: Weinert, Emily E.

In vitro measurement of gas-dependent and redox-sensitive diguanylate 1 cyclase activity

气体依赖性和氧化还原敏感的二鸟苷酸 1 环化酶活性的体外测量

• 发表时间: 2023
• 期刊: Methods in molecular biology
• 作者: Hoque, Nushrat;Helm, Madison;Weinert, Emily E.
• 通讯作者: Weinert, Emily E.