The Ketoreduction and Cyclization of Aromatic Polyketide Biosynthesis

芳香族聚酮生物合成的酮还原和环化

• 批准号: 7827277
• 负责人: Shiou-Chuan Tsai
• 金额: $ 12.63万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7827277
• 关键词: Active Sites; Affect; Affinity; Amino Acid Sequence; Anabolism; Antibiotics; Aromatase; Bicyclo Compounds; Binding; Biological; Biological Factors; CYP19A1 gene; Catalysis; Chemicals; Cloning; Communities; Complement component C1s; Complex; Crystallization; Cyclization; DNA Sequence; Data; Dehydration; Development; Docking; Doxorubicin; Engineering; Environment; Enzyme Interaction; Enzyme Kinetics; Enzymes; Equipment; Estrogen Antagonists; Family; Fatty Acids; Fatty-acid synthase; Figs - dietary; Funding; Gene Cluster; Genes; Grant; Hydro-Lyases; In Vitro; Individual; Investigation; Kinetics; Knowledge; Label; Laboratories; Length; Libraries; Ligands; Link; Macrolide Antibiotics; Modification; Molecular; Multienzyme Complexes; Mutagenesis; Mutate; Mutation; Occupations; Outcome; Parents; Pattern; Peptide Sequence Determination; Pharmacologic Substance; Plicamycin; Population; Positioning Attribute; Principal Investigator; Protein Engineering; Proteins; Public Health; Recovery; Reporting; Research; Research Personnel; Sequence Homology; Simulate; Specificity; Stepparent; Streptomyces; Structure; Structure-Activity Relationship; Substrate Specificity; System; Testing; Tetracyclines; Text; TimeLine; Training; Translating; United States National Institutes of Health; Variant; X-Ray Crystallography; actinorhodin; antitumor agent; base; combinatorial; crosslink; enzyme activity; genetic analysis; graduate student; in vitro Assay; in vivo; innovation; molecular assembly/self assembly; mutant; novel; polyketide synthase; protein complex; protein protein interaction; public health relevance; skills; small molecule; stereochemistry; sugar; tool; tyrosyl-lysine

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.

描述（由申请人提供）：此R01修订申请回答RFA NOT-OD-09-058（通知标题：NIH宣布竞争性修订申请的恢复法案资金可用性）。这是R01授权GM076330的修订申请，题目为“芳香聚酮生物合成的酮还原和环化”，其具体目的是1。确定酮还原酶（KR）的序列-结构-功能关系，使其具有独特的还原特异性。2. 确定芳香化酶/环化酶（ARO/CYC）的序列-结构-功能关系，从而导致其独特的环化特异性。3. 确定蛋白质-蛋白质相互作用在KR和ARO/CYC之间的序列-结构-功能关系中的重要性。我们高兴地报告，我们在实现这三个目标方面的进展已经超过了拟议的时间表。为了进一步加快了解聚酮酮酮还原和环化的研究步伐，我们提出了以下三个目标，它们与母体R01目标不重叠，而是互补的：