Coordinated Mechanistic Approaches to Desulfonation in Two-component FMN Monooxygenases

双组分 FMN 单加氧酶脱磺的协调机制方法

基本信息

批准号：
2105998
负责人：
Holly Ellis
金额：
$ 52.5万
依托单位：
East Carolina University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2105998&HistoricalAwards=false
关键词：
Coordinated Mechanistic Approaches Desulfonation Two

项目摘要

With support from the Chemistry of Life Processes Program in the Division of Chemistry, Professor Holly Ellis and her research team at East Carolina University will study the interesting chemistry carried out by some of the enzymes involved in the acquisition of the element sulfur that bacteria need for survival. A fascinating feature of these enzymes is their ability to change their structures to facilitate the reactions they catalyze. Many of these crucial structural changes and accompanying catalytic events are poorly understood. The two-component enzyme systems selected for this study utilize various structural changes to promote sulfur acquisition. Therefore, these enzyme systems will be ideal to evaluate a multitude of different structural strategies that would be essential for catalysis. The project will also be integrated with the Brody RISE Pre-College Program to train high school students from underrepresented groups in the sciences in different biochemical techniques used to understand enzyme function. An undergraduate student fellowship will be provided in the summer to mentor and encourage participation of underrepresented students in scientific research. Two-component flavin mononucleotide(FMN)-dependent monooxygenases are essential for sulfur acquisition in bacteria when preferred sulfur sources are limiting. These enzymes utilize an FMN reductase and monooxygenase to carry out their critical functional roles; roles that require an elegant coordinated mechanistic strategy for desulfonation. The described objectives will evaluate if the two-component flavin-dependent systems involved in sulfur acquisition have evolved common structural features and mechanistic steps to carry out their complex functional role. Objective 1 is to identify the mechanistic strategies of two-component FMN-dependent monooxygenases that promote desulfonation. Objective 2 is to determine how the structural properties of two-component NAD(P)H:FMN reductases dictate function. Both objectives utilize complementary spectroscopic, kinetic, and biophysical approaches to provide evidence for the desulfonation mechanism and identify oligomeric changes that contribute to flavin transfer in the NAD(P)H:FMN reductases.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.

在化学过程中的化学过程中的支持下，霍莉·埃利斯（Holly Ellis）教授及其在东卡罗来纳大学的研究团队将研究一些涉及获得细菌生存需求的元素硫的酶进行的有趣的化学。这些酶的一个引人入胜的特征是它们改变其结构以促进其催化反应的能力。这些关键的结构变化和随附的催化事件中的许多知识都鲜为人知。为这项研究选择的两组分酶系统利用各种结构变化来促进硫的采集。因此，这些酶系统将是评估多种对催化至关重要的不同结构策略的理想选择。该项目还将与Brody Rise前大学计划集成，以培训来自不同生物化学技术的科学中代表性不足小组的高中生，用于了解酶功能。夏季将提供一项本科生奖学金，以指导并鼓励代表性不足的学生参与科学研究。当首选的硫源被限制时，两种成分黄素单核苷酸（FMN）依赖性单加氧酶对于细菌中的硫是必不可少的。这些酶利用FMN还原酶和单加氧酶来扮演关键功能。需要优雅的脱硫机械策略的角色。所描述的目标将评估涉及硫磺采集的两分量黄素依赖性系统是否发展了共同的结构特征和机械步骤，以执行其复杂的功能。目的1是确定促进脱硫化的两个组分依赖性单加氧酶的机械策略。目标2是确定两个组分NAD（P）H：FMN还原酶的结构特性如何决定功能。这两种目标都利用互补的光谱，动力学和生物物理方法来提供脱硫的机制的证据，并确定有助于在NAD（P）H：FMN还原酶转移黄素转移的寡头变化。此奖项。该奖项反映了NSF的法定任务，反映了通过评估的支持者，其知识范围的范围和众所周知的范围众所周知。