METABOLISM AND FUNCTION OF CYCLIC NUCLEOTIDES

拟议研究的总体目标是定义分子 正常人环核苷酸代谢调控机制的研究进展 哺乳动物组织。具体重点将放在 的鉴定、特征、调节和生理作用 CGMP刺激和钙/钙调蛋白依赖家族的每个成员 环核苷酸磷酸二酯酶。四个主要方法将是 强调。首先，每个不同同工酶的全长cDNA将是 分离、鉴定和表达。第二，领域 组织和互动将使用诱变技术进行解剖， 缺失分析和嵌合PDE构建。第三，生理学 通过磷酸化和激素的功能和调节 在完整细胞和体外实验中都进行了研究。最后，两项前景看好的新技术 新型环核苷酸的cDNA分离技术 将探索磷酸二酯酶。

The general aims of the proposed research are to define the molecular mechanisms for the regulation of cyclic nucleotide metabolism in normal mammalian tissues. Specific emphasis will be placed on the identification, characterization, regulation and physiological roles for each member of the cGMP-stimulated and Ca++/calmodulin-dependent families of cyclic nucleotide phosphodiesterase. Four major approaches will be emphasized. First, full length cDNAs for each different isozyme will be isolated, characterized and expressed. Secondly, the domain organizations and interactions will be dissected using mutagenesis, deletion analysis, and chimeric PDE constructs. Thirdly, physiological function and regulation by phosphorylation and by hormones will be studied in both intact cells and in in vitro. Finally, two promising new techniques for isolation of cDNAs for novel cyclic nucleotide phosphodiesterases will be explored.