STRUCTURE AND FUNCTION OF BIOSYNTHETIC ENZYMES

生物合成酶的结构和功能

基本信息

批准号：
7472416
负责人：
DAVID W CHRISTIANSON
金额：
$ 28.08万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-08-01 至 2010-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7472416
关键词：
Accounting Active Sites Address Ally Amino Acid Sequence Amino Acid Substitution Anabolism Analgesics Antifungal Antibiotics Binding Biological Biological Assay Biological Factors Biology Bond-It C10 Canes Carbon Catalysis Chemicals Chemistry Chemotherapy-Oncologic Procedure Class Commit Complex Crystallization Cyclization Diphosphates Diterpenes Drug Delivery Systems Engineering Enzymes Evolution Family Foundations Funding Generations Goals Health Hot Spot Human Knowledge Magnesium Mass Fragmentography Metabolic Pathway Metabolism Metals Methylation Methyltransferase Modification Molecular Conformation Mutate Mycoses Natural Products Chemistry Nature Organism Paclitaxel Pharmaceutical Preparations Play Preparation Property Protons Reaction Reporting Roentgen Rays Role Site Solid Staging Sterol Biosynthesis Pathway Sterols Structure Structure-Activity Relationship Taxus Terpenes Terpenoid Biosynthesis Pathway Time Triterpenes Variant Work X-Ray Crystallography analog aqueous aristolochene aristolochene synthase base biosynthetic product chemical bond chemical reaction cholesterol biosynthesis combinatorial chemistry comparative enzyme structure farnesyl pyrophosphate fascinate fungus geranylgeranyl diphosphate monoterpene cyclase novel stereochemistry structural biology taxadiene synthase three dimensional structure trichodiene synthase

项目摘要

DESCRIPTION (provided by applicant): Thousands of terpenes and terpenoid derivatives found throughout Nature are involved in diverse biosynthetic and metabolic pathways such as cholesterol biosynthesis in humans and paclitaxel (Taxol) synthesis in the Pacific yew. Notably, many terpenoids have been used as medicinal agents since times of antiquity due to their analgesic, antibiotic, and antifungal properties. In spite of the universal importance of this family of natural products for human health, it is remarkable that the three-dimensional structures of terpenoid cyclases have only been reported relatively recently. Terpenoid cyclases (a.k.a. synthases) catalyze the specific cyclization of a common allylic pyrophosphate substrate, such as farnesyl diphosphate, into one of hundreds of possible products. The terpenoid cyclase plays a critical role as a template in "channeling" the precise substrate and intermediate conformations leading to the formation of one exclusive product. Thus, the terpenoid cyclases comprise an exciting class of biosynthetic enzymes from both the biological and the chemical perspectives. In the current funding period, we have determined the first X-ray crystal structure of a monoterpene cyclase, (+)- bornyl diphosphate synthase; we have determined the fifth crystal structure of a sesquiterpene cyclase, aristolochene synthase from A.terreus; and we have established the structural basis for aberrant product formation by site-specific variants of trichodiene synthase. We aim to build upon this outstanding foundation in the next funding period by dissecting detailed structure-biosynthetic diversity relationships in trichodiene synthase. Specific!ally, we will study site-specific variants with altered metal binding properties, and we will also study variants engineered to generate alternative products. We will also study the structural basis for the evolution and fidelity of aristolochene synthase from A. terreus and P. roqueforti. In order to broaden our knowledge of structure-function relationships in the greater family of terpenoid cyclases, we will also study the diterpene cyclase taxadiene synthase. Finally, we will determine the X-ray crystal structure of sterol methyltransferase, a potential drug target for the treatment of fungal infections. Now that we have established a solid foundation in the study of biosynthetic enzymes that generate cyclic terpene products, we will now study an enzyme that utilizes a cyclic terpene substrate in a novel chemical reaction that further diversifies the biosynthetic array of cyclic terpene natural products.

描述（由申请人提供）：自然界中发现的数千种萜烯和萜类衍生物参与多种生物合成和代谢途径，例如人类的胆固醇生物合成和太平洋红豆杉的紫杉醇（紫杉醇）合成。值得注意的是，许多萜类化合物由于其镇痛、抗生素和抗真菌特性，自古以来就被用作药物。尽管这一天然产物家族对人类健康具有普遍重要性，但值得注意的是，萜类环化酶的三维结构直到最近才被报道。萜类环化酶（又名合酶）催化常见烯丙基焦磷酸底物（例如法呢基二磷酸）的特异性环化，形成数百种可能的产物之一。萜类环化酶作为模板在“引导”精确底物和中间构象从而导致一种独特产物的形成中发挥着关键作用。因此，从生物学和化学角度来看，萜类环化酶包含一类令人兴奋的生物合成酶。在本期资助期间，我们确定了第一个单萜环化酶（+）-冰片基二磷酸合酶的X射线晶体结构；我们已经确定了来自 A.terreus 的倍半萜环化酶、马兜铃烯合酶的第五种晶体结构；我们已经建立了单端孢二烯合酶位点特异性变体形成异常产物的结构基础。我们的目标是在下一个资助期通过剖析单族二烯合酶的详细结构-生物合成多样性关系来建立这一杰出的基础。具体来说，我们将研究具有改变的金属结合特性的位点特异性变体，我们还将研究设计用于生成替代产品的变体。我们还将研究来自 A. terreus 和 P. roqueforti 的马兜铃烯合酶的进化和保真度的结构基础。为了拓宽我们对萜类环化酶大家族的结构-功能关系的了解，我们还将研究二萜环化酶紫杉二烯合酶。最后，我们将确定甾醇甲基转移酶的 X 射线晶体结构，甾醇甲基转移酶是治疗真菌感染的潜在药物靶点。现在我们已经在产生环状萜烯产物的生物合成酶的研究中奠定了坚实的基础，我们现在将研究一种在新型化学反应中利用环状萜烯底物的酶，进一步使环状萜烯天然产物的生物合成系列多样化。