ISOPRENOID COMPOUNDS FOR BIOLOGICAL STUDIES

类异戊二烯化合物在所有生物体中都有合成，它们在新陈代谢、细胞结构以及细胞内和细胞间的信号传递中起着重要作用。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进行酶分析、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以在商业上使用。生物学家和生物化学家除了需要最常见的衍生物或特殊的标记模式外，其他都需要，他们必须求助于昂贵的“定制”合成。带有荧光或光亲和标签的分子，除了少数学术实验室外，并非所有实验室都能找到。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)开发了纯化和储存类异戊二烯二磷酸异构酶和法尼酯二磷酸合成酶的方法，以及(c)合成了由附加在类异戊二烯二磷酸上的光亲和性二苯甲酮组成的探针。在II期，我们将扩大我们的产品线，包括更广泛的天然存在的非市售类异戊二烯分子，添加光亲和标签，并添加一系列荧光化合物用于测定，结合测量和标记戊烯化蛋白。一类类异戊二烯酶，包括二磷酸异戊二烯异构酶；一组经过工程改造的戊烯基转移酶，可从简单的C5构建块合成香叶基（C10）、香叶基（C15）、香叶基香叶基（C20）和香叶基香叶基（C25）二磷酸；和蛋白法尼基转移酶，将单独提供或作为试剂盒用于合成标记分子。拟议的商业应用：类异戊二烯化合物越来越多地用于药物发现分析、动脉粥样硬化和癌症领域、研究细胞信号机制和细胞代谢研究。ERL的类异戊二烯系列产品将为研究人员提供促进这些研究的化学物质和酶。

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.