ISOPRENOID COMPOUNDS FOR BIOLOGICAL STUDIES

用于生物学研究的异戊二烯化合物

• 批准号: 6381257
• 负责人: CHARLES DALE POULTER
• 金额: $ 24.69万
• 依托单位: ECHELON BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 2003-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6381257
• 关键词: alkyltransferase; biotechnology; chemical synthesis; enzyme activity; fluorescent dye /probe; isoprenoid; peptide chemical synthesis; photochemistry; protein purification

Isoprenoid compounds are synthesized in all living organisms where they play essential roles in metabolism, cellular architecture, and signalling both within and between cells. The molecules required for the fundamental building reactions in the pathway are needed in unlabeled, stable isotope labeled, and radiolabeled form for enzyme assays, drug screening protocols, metabolism studies, and cell biology experiments. Only a limited number of these compounds are available commercially. Biologists and biochemists who require all but the most common derivatives or special labeling patterns must resort to costly "custom" syntheses. Molecules with fluorescent or photoaffinity tags are not available in all but a few academic laboratories. In phase I, (a) we optimized the "large scale" synthesis of simple linear C5, C10, C15, and C20 isoprenoid diphosphates, (b)developed procedures for purification and storage of isopentenyl diphosphate isomerase and farnesyl diphosphate synthase, and (c) synthesized photoaffinity probes consisting of photolabile benzophenone moieties appended to isoprenoid diphosphates. In phase II we will expand our product line to include a wider range of naturally occurring isoprenoid molecules not commercially available, add photoaffinity labels, and add a family of fluorescent compounds for assays, binding measurements, and tags for prenylated proteins. A set of isoprenoid enzymes, including isopentenyl diphosphate, isomerase; an engineered set of prenyltransferases selective for synthesis of geranyl (C10), farnesyl (C15), geranylgeranyl (C20), and farnesylgeranyl (C25) diphosphates from simple C5 building blocks; and protein farnesyltransferase, will be provided individually or as kits for synthesis of labeled molecules. PROPOSED COMMERCIAL APPLICATIONS: Isoprenoid compounds are increasingly used in drug discovery assays, in the areas of atherosclerosis and cancer, to study cell signalling mechanisms, and studies of cellular metabolism. ERL's line of isoprenoid products will provide researchers with chemicals and enzymes to facilitate these studies.

类异戊二烯化合物在所有生物体中合成，它们在细胞内和细胞间的代谢、细胞结构和信号传导中发挥重要作用。该途径中基本构建反应所需的分子需要以未标记、稳定同位素标记和放射性标记的形式用于酶测定、药物筛选方案、代谢研究和细胞生物学实验。只有有限数量的这些化合物可商购获得。除了最常见的衍生物或特殊的标记模式之外，生物学家和生物化学家需要所有的衍生物或特殊的标记模式，他们必须求助于昂贵的“定制”合成。除了少数学术实验室外，并非所有实验室都能获得带有荧光或光亲和标记的分子。在第一阶段，（a）我们优化了简单线性C5，C10，C15和C20类异戊二烯二磷酸的“大规模”合成，（B）开发了异戊烯基二磷酸异构酶和法呢基二磷酸合酶的纯化和储存程序，和（c）合成了由附加到类异戊二烯二磷酸的光不稳定二苯甲酮部分组成的光亲和探针。在第二阶段，我们将扩大我们的产品线，包括更广泛的天然存在的类异戊二烯分子没有商业上，添加光亲和标记，并添加一个家庭的荧光化合物的测定，结合测量，和标签的异戊二烯化蛋白质。将单独提供一组类异戊二烯酶，包括异戊烯基二磷酸异构酶;一组工程化的异戊二烯基转移酶，其对从简单的C5结构单元合成香叶基（C10）、法呢基（C15）、香叶基香叶基（C20）和法呢基香叶基（C25）二磷酸具有选择性;以及蛋白质法呢基转移酶，或作为试剂盒提供用于合成标记的分子。拟议的商业应用：类异戊二烯化合物越来越多地用于动脉粥样硬化和癌症领域的药物发现测定，以研究细胞信号传导机制和细胞代谢的研究。ERL的类异戊二烯产品线将为研究人员提供化学品和酶，以促进这些研究。