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）：Polyketides天然产品已表现出庞大的药理特性，导致150亿美元的行业1,2。这些多功能化合物的生物合成机制尚不清楚。通过探索和理解导致聚酮化合物生物合成的机制，我们希望通过体外重组聚酮化合酶合酶（PKS）有效合成大量的desired polyketide药物。我们的长期目标是生物工程聚酮化合酶以创建新的治疗剂。至少有三种类型的PKS，但是该提案的重点是II型PKS，由五到十种酶组成，这些酶进行链条伸长和修改。该应用的总体目的是提供对II型PKS的芳香化酶/环化酶（ARO/CYC）的结构见解。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，我们的目的研究与NIH的使命有关，该使命与最终保护和改善健康的创新方法有关。在我们的初步数据的指导下，该假设将通过两个特定目的检验：1）确定聚酮化合物二域芳香蛋白酶/旋风酶/旋风酶（ARO/CYC）的晶体结构； 2）工程Di域ARO/CYC以合理控制第一圈环化特异性。拟议的研究很重要，因为有必要确定II型PKS中二域ARO/CYC的环化特异性的分子基础。到目前为止，这种知识差距已经阻止了具有新的环化模式的新芳族聚酮化合物的发展。