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

有了这个奖项，化学部的生命过程化学项目资助了弗吉尼亚理工大学的Pablo Sobrado博士和John。来自密苏里大学的J. Tanner研究了一种被称为黄素依赖n-单加氧酶（NMOs）的酶的结构和化学。NMOs在微生物如何为各种目的制造大量天然化合物方面发挥着不可或缺的作用。这些化合物还具有有益社会的生物活性。虽然许多依赖黄素的酶是众所周知的，并且已经被彻底地表征，但NMOs是在家族树中相对较新发现的分支，并且在常见的方面与其他成员不相似。这导致了一种假设，即新的有趣的结构和化学物质可能在nmo催化的反应中起作用。研究者选择了几个nmo的例子进行深入的研究，以比较和对比他们的行为。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。提出的实验方法将为现代酶学和生物物理化学的本科生和研究生的培训提供一个平台。各实验室亦致力推行策略，以提高科研人员的多样性。该项目通过新的n -单加氧酶的结构和生化表征扩展了我们对黄酶多样性的认识。由于这些酶与蛋白质数据库中已有的蛋白质相似性较低，因此这些结构有望表现出新的特征，并揭示与其他酶的意想不到的关系。这进一步推进了我们对这些酶的进化关系的理解。通过表征“双羟基化”NMOs，探索B族NMOs中新发现的黄素“扑动”运动，以及在晶体中捕获迄今为止难以捉摸的黄酶配体复合物以确定结构，解决了我们对黄酶机制理解中的关键知识空白。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.