Collaborative Research: Structure and function of flavin-dependent N-monooxygenases

合作研究：黄素依赖性 N-单加氧酶的结构和功能

基本信息

批准号：
2003658
负责人：
Pablo Sobrado
金额：
$ 33万
依托单位：
Virginia Polytechnic Institute and State 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：
2020-07-01 至 2024-05-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2003658&HistoricalAwards=false
关键词：
Collaborative Research Structure function flavin

项目摘要

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Pablo Sobrado from Virginia Tech and John. J. Tanner from the University of Missouri to investigate the structure and chemistry of enzymes known as flavin-dependent-N-monooxygenases (NMOs). NMOs play integral parts in how microorganisms make numerous natural compounds for a variety of purposes. These compounds can also have useful bioactivities that benefit society. While many flavin-dependent enzymes are well known and have been thoroughly characterized, the NMOs are a relatively recently discovered branch on the family tree, and do not resemble the other members in common ways. This leads to the hypothesis that new and interesting structures and chemistries are likely to be at play in the reactions catalyzed by the NMOs. The investigators choose several examples of NMOs for in depth examination in order to compare and contrast their behaviors. Further, a highly multidisciplinary approach is proposed to provide the broadest possible understanding of the enzymes. The proposed experimental approach will provide a platform for the training of undergraduate and graduate students in modern enzymology and biophysical chemistry. The laboratories are also committed to strategies to enhance diversity in the scientific research workforce.This project expands our knowledge of the diversity of flavoenzymes through the structural and biochemical characterization of new N-monooxygenases. Because these enzymes have low similarity to proteins already in the Protein Data Bank, the structures are expected to exhibit novel features and reveal unanticipated relationships to other enzymes. This further advances our understanding of the evolutionary relatedness of these enzymes. Also addressed are key knowledge gaps in our understanding of flavoenzyme mechanisms by characterizing “double-hydroxylating” NMOs, exploring a newly discovered flavin “flapping” motion in Group B NMOs, and trapping heretofore-elusive flavoenzyme-ligand complexes in crystallo for structure determination.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.

凭借该奖项，化学部的生命过程化学项目正在资助弗吉尼亚理工大学的 Pablo Sobrado 博士和密苏里大学的 John J. Tanner 研究黄素依赖性 N- 酶的结构和化学。单加氧酶 (NMO) 在微生物生产多种天然化合物的过程中发挥着不可或缺的作用，这些化合物也具有造福社会的有用生物活性。尽管酶是众所周知的并且已经被彻底表征，但 NMO 是家族树上相对较新发现的分支，并且与其他成员在常见方面并不相似，这导致了这样的假设：新的、有趣的结构和化学性质可能是存在的。研究人员选择了几个 NMO 的例子进行深入研究，以比较和对比它们的行为。此外，提出了一种高度多学科的方法，以提供对酶的最广泛的了解。将为本科生和研究生提供现代酶学和生物物理化学方面的培训平台。实验室还致力于增强科研队伍多样性的战略。该项目通过结构和生物化学扩展了我们对黄素酶多样性的了解。由于这些酶与蛋白质数据库中已有的蛋白质相似性较低，因此这些酶的结构有望表现出新颖的特征，并揭示与其他酶的意想不到的关系，这进一步加深了我们对这些酶的理解。通过表征“双羟基化”NMO、探索 B 组 NMO 中新发现的黄素“扑动”运动以及捕获迄今为止难以捉摸的黄素酶配体，还解决了我们对黄素酶机制理解中的关键知识空白。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进行评估，被认为值得支持标准。