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计划正在资助弗吉尼亚理工大学的巴勃罗索布拉多博士和约翰。密苏里州大学的J.坦纳研究称为黄素依赖性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许多天然化合物方面发挥着不可或缺的作用。 这些化合物还可以具有有益于社会的有用的生物活性。 虽然许多黄素依赖性酶是众所周知的，并已被彻底表征，但NMO是家族树上相对较新发现的分支，并且在常见方式上与其他成员不相似。 这导致了一种假设，即新的和有趣的结构和化学可能在NMOs催化的反应中起作用。 研究者选择了几个非军事组织的例子进行深入研究，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，以提供最广泛的酶的理解。 建议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。实验室还致力于提高科学研究队伍的多样性的战略。该项目通过新的N-单加氧酶的结构和生物化学特性扩展了我们对黄素酶多样性的认识。由于这些酶与蛋白质数据库中已有的蛋白质具有低相似性，因此预计这些结构将表现出新的特征，并揭示与其他酶的意外关系。 这进一步推进了我们对这些酶的进化相关性的理解。 还解决了我们理解黄素酶机制的关键知识差距，通过表征“双羟基化”NMOs，探索B组NMOs中新发现的黄素“拍打”运动，并捕获了迄今为止难以捉摸的黄素酶该奖项反映了NSF的法定使命，并已被认为是值得通过评估使用基金会的智力价值和更广泛的影响力的支持审查标准。

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.