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

通过这一奖项，化学部的生命过程化学项目资助了弗吉尼亚理工大学和约翰大学的巴勃罗·索布拉多博士。来自密苏里大学的J.Tanner研究被称为黄素依赖N-单加氧酶(NMOS)的酶的结构和化学。NMOS在微生物为各种目的制造许多天然化合物的过程中发挥着不可或缺的作用。这些化合物还可以具有有益的生物活性，造福社会。虽然许多黄素依赖的酶是众所周知的，并且已经得到了彻底的表征，但NMOS是家谱上相对较新发现的一个分支，在共同的方面与其他成员并不相似。这导致了一种假设，即新的和有趣的结构和化学可能在NMOS催化的反应中发挥作用。调查人员选择了几个NMOS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，以提供对酶的最广泛的了解。建议的实验方法将为培养现代酶学和生物物理化学的本科生和研究生提供一个平台。这些实验室还致力于加强科学研究工作的多样性的策略。这个项目通过对新的N-单加氧酶的结构和生化特性的研究，扩大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，预计这些结构将显示出新的特征，并揭示与其他酶的意想不到的关系。这进一步促进了我们对这些酶的进化相关性的理解。本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智力优势和更广泛的影响审查标准对黄素酶的评估标准进行了评估。

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.