The Ketoreduction and Cyclization of Aromatic Polyketide Biosynthesis

描述（由申请人提供）：此R 01修订申请回答RFA NOT-OD-09-058（通知标题：NIH宣布恢复法案资金可用于竞争性修订申请）。这是R 01资助GM 076330的修订申请，标题为“芳香聚酮生物合成的酮还原和环化”，其具体目标是1。确定酮还原酶（KR）的序列-结构-功能关系，导致其独特的还原特异性。2.确定芳香化酶/环化酶（ARO/CYC）的序列-结构-功能关系，导致其独特的环化特异性。3.确定蛋白质-蛋白质相互作用对KR和ARO/CYC之间的序列-结构-功能关系的重要性。我们高兴地报告，我们在实现这三个目标方面的进展超过了拟议的时间表。为了进一步加快研究步伐，了解聚酮化合物酮还原和环化，我们提出了以下三个目标，这些目标不重叠，而是对母体R 01目标的补充： AIM 1.使用化学探针确定酮还原的分子基础。 AIM 2.用化学探针确定芳香聚酮环化反应的分子基础。 AIM 3.解决化学交联的多结构域PKS复合物的晶体结构。这项研究意义重大，因为其结果将回答有关如何精确控制聚酮化合物还原和环化的重要问题。它也是创新的，提供了新的信息KR和ARO/CYC在分子水平上以前没有实现。长期的生物医学相关性是，聚酮化合物研究界可以应用从该提议确定的序列-结构-功能关系，通过蛋白质工程使“非天然”天然产物的群体多样化，从而可以产生具有不同酮还原和环化模式的新型聚酮化合物库。此次修订将加快聚酮合酶（PKS）的科学研究克里思，并允许为三名研究生创造和保留就业机会，雇用一名新的博士后研究人员，并采购实验室设备以加快拟议研究的克里思。 公共卫生相关性：预计这将对公众健康产生积极影响，因为它将允许开发新的“非天然”天然产品，这些产品可以筛选新的药物活性。与此同时，在该提案中获得的关于将PKS序列结构与聚酮酮还原和环化过程中的催化、底物特异性和蛋白质-蛋白质相互作用相关联的基础新知识预计将对天然产物研究界产生重大影响。

DESCRIPTION (provided by applicant):This R01 revision application answers RFA NOT-OD-09-058 (Notice Title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications). This is a revision application for the R01 grant GM076330, titled "The Ketoreduction and Cyclization of Aromatic Polyketide Biosynthesis", whose specific aims were 1. Determine the sequence-structure-function relationship of ketoreductase (KR) that leads to its unique reduction specificity. 2. Determine the sequence-structure-function relationship of aromatase/cyclase (ARO/CYC) that leads to its unique cyclization specificity. 3. Determine the importance of protein-protein interactions on the sequence-structure-function relationship between KR and ARO/CYC. We were pleased to report that our progress towards these three aims has exceeded the proposed timelines. In order to further accelerate the research pace towards understanding polyketide ketoreduction and cyclization, we proposed the following three aims that do not overlap, but rather complementary to the parent R01 aims: AIM 1. Determine the molecular basis of ketoreduction using chemical probes. AIM 2. Determine the molecular basis of aromatic polyketide cyclization using chemical probes. AIM 3. Solve crystal structures of chemically cross-linked multi-domain PKS complexes. The proposed research is significant, because the outcome will answer important questions about how polyketide reduction and cyclization are precisely controlled. It is also innovative by providing new information about KR and ARO/CYC at a molecular level not achieved previously. The long-term biomedical relevance is that the polyketide research community can apply the sequence-structure-function relationships determined from this proposal to diversify the population of "unnatural" natural products via protein engineering, such that a library of novel polyketides with different ketoreduction and cyclization patterns can be produced. The revision will accelerate the tempo of scientific research on polyketide synthase (PKS) and allow for job creation and retention for three graduate students and the hiring of one new postdoctoral researcher, and procuring laboratory equipments to accelerate the tempo of the proposed research. PUBLIC HEALTH RELEVANCE: This is expected to positively affect public health, because it will allow the development of new "unnatural" natural products that can be screened for new pharmaceutical activities. Meanwhile, the fundamental new knowledge obtained in this proposal on correlating PKS sequence-structure with the catalysis, substrate specificity and protein-protein interactions during polyketide ketoreduction and cyclization is expected to have a high impact on the natural product research communities.