Elucidating the Intricacies of Type II Aromatase/Cyclase Domains

阐明 II 型芳香酶/环化酶结构域的复杂性

• 批准号: 8257727
• 负责人: Grace Marie Festin Caldara
• 金额: $ 4.22万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8257727
• 关键词: Anabolism; Antibiotics; Architecture; Aromatase; Biological Factors; Biomedical Engineering; CYP19A1 gene; Chemical Structure; Complex; Consumption; Crystallography; Cyclization; Data; Development; Engineering; Enzymes; Equipment; Exhibits; Genetic Recombination; Goals; Health; High Pressure Liquid Chromatography; Home environment; Human; In Vitro; Individual; Industry; Knowledge; Laboratories; Lactones; Lead; Mass Spectrum Analysis; Mission; Modification; Molecular; Multienzyme Complexes; Mutagenesis; Mutation; NMR Spectroscopy; Organ Transplantation; Outcome; Pathway interactions; Pattern; Pharmaceutical Preparations; Pharmacologic Substance; Play; Polyenes; Production; Property; Public Health; Reproduction; Research; Resolution; Role; Solutions; Source; Specificity; Structure; Synchrotrons; System; Tertiary Protein Structure; Testing; Therapeutic; Time; Work; X-Ray Crystallography; base; cancer therapy; improved; innovation; insight; novel therapeutics; polyketide synthase; prevent; protein protein interaction; tool

DESCRIPTION (provided by applicant): Polyketides natural products have exhibited vast pharmacological properties resulting in a $15 billion dollars industry 1,2. The biosynthetic mechanism of these versatile compounds is not well understood. By exploring and understanding the mechanisms that lead to polyketide biosynthesis, we hope to eventually bioengineer new polyketide therapeutics through in vitro recombination of polyketide synthase (PKS) to efficiently synthesize large quantities of the desired polyketide drug. Our long-term goal is to bioengineer polyketide enzymes to create new therapeutics. There are at least three types of PKSs, but the focus of this proposal is type II PKSs, which consist of five to ten enzymes that conduct chain elongations and modifications iteratively. The overall objective of this application is to provide structural insight into the aromatase/cyclase (ARO/CYC) of type II PKSs. Our central hypothesis is that we can apply structure-based mutagenesis to change the product outcome. Thus our purposed research is relevant to the NIH's mission that pertains to an innovative approach for ultimately protecting and improving health. Guided by our preliminary data, the hypothesis will be tested by two specific aims: 1) Determining the crystal structures of the polyketide di-domain aromatase/cyclase (ARO/CYC); and 2) Engineering the di-domain ARO/CYC to rationally control first-ring cyclization specificity. The proposed studies are significant, because there is a need to determine the molecular basis of cyclization specificities by di-domain ARO/CYCs in type II PKSs. This knowledge gap has so far prevented the development of new aromatic polyketides with new cyclization patterns.

描述（由申请人提供）：聚酮化合物天然产品表现出巨大的药理学特性，导致其产业价值达 150 亿美元1,2。这些多功能化合物的生物合成机制尚不清楚。通过探索和了解聚酮化合物生物合成的机制，我们希望最终通过聚酮化合物合成酶（PKS）的体外重组来生物工程设计新的聚酮化合物疗法，以有效地合成大量所需的聚酮化合物药物。我们的长期目标是对聚酮酶进行生物工程以创造新的疗法。 PKS 至少有三种类型，但该提案的重点是 II 型 PKS，它由五到十种反复进行链延伸和修饰的酶组成。该应用的总体目标是深入了解 II 型 PKS 的芳香酶/环化酶 (ARO/CYC) 的结构。我们的中心假设是我们可以应用基于结构的诱变来改变产品结果。因此，我们有针对性的研究与 NIH 的使命相关，该使命涉及最终保护和改善健康的创新方法。在我们的初步数据的指导下，该假设将通过两个具体目标进行检验：1）确定聚酮化合物双结构域芳香酶/环化酶（ARO/CYC）的晶体结构； 2）设计双结构域ARO/CYC以合理控制第一环环化特异性。拟议的研究意义重大，因为需要确定 II 型 PKS 中双结构域 ARO/CYC 环化特异性的分子基础。迄今为止，这种知识差距阻碍了具有新环化模式的新型芳香族聚酮化合物的开发。