REGULATION OF CYCLIC NUCLEOTIDE FORMATION AND ACTION

调查员的长期目标是了解准确的 正常人环核苷酸合成的调控及其功能 组织和各种疾病状态。建议的工作将强调 环状GMP的调控和作用。将开发模型系统以 检测荷尔蒙和大肠杆菌耐热性肠毒素的调节 两种完整细胞中鸟苷环化酶的活性和GMP的循环积累 和无细胞系统。E.Coli热稳定性的偶联机制 肠毒素与环化GMP合成肠黏膜的结合 内皮依赖性血管扩张剂对血管平滑肌的作用 环状GMP合成将被用作两个模型系统的例子 鸟苷环化酶调节。此外，还将进行实验以 在无细胞系统中重建鸟苷环化酶的激素调节 使用模型细胞培养系和组织。此外，红血球幽灵 会负载各种与环核苷酸有关的大分子 合成、水解和功能。这些被加载的幽灵将被融合到 修饰环核苷酸代谢的细胞系和感兴趣的组织 和职能，以便评估它们在各种进程和/或 磷酸化事件。因此，将采取几种方法 同时提供新的技术和信息来检查 环核苷酸的形成和作用的调节。

The long term objectives of the investigator are to understand the precise regulation of cyclic nucleotide synthesis and their function in normal tissues and various disease states. The work proposed will emphasis the regulation and role of cyclic GMP. Model systems will be developed to examine hormonal and E. Coli heat-stable enterotoxin regulation of guanylate cyclase activity and cyclic GMP accumulation in both intact cell and cell-free systems. The mechanisms of coupling of E. Coli heat-stable enterotoxin binding with cyclic GMP synthesis intestinal mucosa and the effects of endothelial dependent vasodilators on vascular smooth muscle cyclic GMP synthesis will be used as example of two model systems for guanylate cyclase regulation. Furthermore experiments will be conducted to reconstitute hormonal regulation of guanylate cyclase in cell-free systems using model cell culture lines and tissues. In addition red cell ghosts will be loaded with various macromolecules involved in cyclic nucleotide synthesis, hydrolysis and function. These loaded ghosts will be fused to cell lines and tissues of interest to modify cyclic nucleotide metabolism and function in order to assess their role in various processes and/or phosphorylation events. Thus, several approaches will be conducted simultaneously to provide new techniques and information to examine the regulation of cyclic nucleotide formation and action.