Enzymatic formation of the hydrocarbon skeletons underlying diterpenoid diversity

二萜多样性背后的碳氢化合物骨架的酶促形成

• 批准号: 8706175
• 负责人: REUBEN JOHN PETERS
• 金额: $ 29.49万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8706175
• 关键词: Adenylate Cyclase; Anabolism; Binding; Biochemical; Biological; Biological Factors; Carbon; Commit; Complex; Cyclization; Diphosphates; Diterpenes; Drug Targeting; Electrostatics; Engineering; Environment; Enzymes; Family; Forskolin; Goals; Grant; Human; Hydrocarbons; Immune response; Individual; Infection; Investigation; Isomerism; Lead; Libraries; Malignant Neoplasms; Mediating; Medicine; Metabolic Pathway; Microbe; Mycobacterium tuberculosis; Organism; Outcome; Paclitaxel; Pharmacologic Substance; Phorbol Esters; Plants; Play; Production; Protein Kinase C; Reaction; Research; Role; Skeleton; Source; Structure; Terpenes; Vertebral column; Work; base; geranylgeranyl diphosphate; insight; isoprenoid; macromolecule; novel; pathogen; prostratin; protonation; skeletal; terpene synthase; tool; tumor

DESCRIPTION (provided by applicant): Natural products have a proven record of providing a significant fraction, either directly or as lead compounds, of human medicines. Among natural products, the terpenoids (isoprenoids) stand out as being the largest class (>50,000 already known), with the 20-carbon diterpenoids targeted here forming a significant fraction of these (>12,000 known). Indeed, the extensive diversification of diterpenoids indicates that the manifold structures that can be formed from this C20 backbone provide a rich source of biological activity, and a number of these natural products are used as pharmaceuticals (e.g., the anti- cancer paclitaxel/TaxolTM and anti-biotic mutilins) or research tools (e.g., the protein kinase C activating and tumor promoting phorbol esters, and adenylate cyclase activator forskolin). Conversely, we have recently discovered that the human pathogen Mycobacterium tuberculosis utilizes a diterpenoid, isotuberculosinol, to modulate our initial immune response to infection. Accordingly, we propose here to continue our productive investigations of the enzymes, from both plants and microbes, that catalyze formation of the underlying hydrocarbon skeletal structures. Specifically, we will build on our findings from the previous grant period, which includes not only the discovery of isotuberculosinol, but elucidation of some of the first diterpene synthase crystal structures as well. This proposal then advances our long- term goal of engineering enzymes and metabolic pathways for the production of targeted libraries and specific individual terpenoid 'natural' products for pharmaceutical investigation and use. In particular, the objective of this proposal is a detailed structure-function investigation of the synthases/cyclases that catalyze formation of the hydrocarbon skeletons underlying the structural diversity of diterpenoid natural products. These enzymes catalyze complex electrophilic reactions that form new carbon-carbon bonds, and play important roles in initiating diterpenoid biosynthesis. Accordingly, our studies have implications not only for providing increased access to such natural products, but also insights into potential drug targets, specifically in the widespread pathogen M. tuberculosis.

描述（由申请人提供）：天然产品具有直接或作为人类药物的先导化合物提供重要部分的证明记录。在天然产物中，萜类化合物（类异戊二烯）是最大的一类（已知的> - 5万），其中20碳二萜类化合物占很大一部分（已知的> - 12万）。事实上，二萜的广泛多样化表明，由C20骨架形成的多种结构提供了丰富的生物活性来源，许多这些天然产物被用作药物（例如抗癌紫杉醇/TaxolTM和抗生素多重素）或研究工具（例如蛋白激酶C激活和促进肿瘤的佛波酯，以及腺苷酸环化酶激活剂forskolin）。相反，我们最近发现，人类病原体结核分枝杆菌利用二萜异结核醇来调节我们对感染的初始免疫反应。因此，我们建议在这里继续我们的酶的生产研究，从植物和微生物，催化形成潜在的碳氢化合物骨架结构。具体地说，我们将建立在我们之前的研究成果的基础上，这些发现不仅包括异结核菌醇的发现，还包括对一些第一二萜合成酶晶体结构的阐明。该提案推进了我们的长期目标，即工程酶和代谢途径，以生产目标文库和特定的个体萜类“天然”产品，用于药物研究和使用。特别地，本提案的目的是对催化碳氢化合物骨架形成的合成酶/环化酶进行详细的结构-功能研究，这些合成酶/环化酶是二萜天然产物结构多样性的基础。这些酶催化复杂的亲电反应，形成新的碳-碳键，并在启动二萜生物合成中起重要作用。因此，我们的研究不仅对增加获得这种天然产物的途径有意义，而且对潜在的药物靶点，特别是在广泛传播的病原体结核分枝杆菌中也有意义。