REGULATION OF CYCLIC NUCLEOTIDE FORMATION AND ACTION

本提案的具体目的是确定 激素、类固醇和药物调节的机制 鸟苷酸环化酶和环GMP合成的同工酶形式。 激素受体偶联的精确机制， 不同的鸟苷酸环化酶同工酶将在 相当详细 实验将利用完整的细胞培养物 和组织以及粗品和纯化成分及其 在无细胞制剂中重构。 心房的影响 利钠因子和E.大肠杆菌耐热肠毒素 GMP合成以及其他配体和激素将作为 模型和原型。 ATP的作用， 其他核苷酸和最近纯化的蛋白质激活剂， 还将检查鸟苷酸环化酶。 这些的作用 材料以及蛋白质磷酸化， 鸟苷酸环化酶调节和激素中的去磷酸化 将检查与环GMP合成的偶联。 方法将 包括偶联因子的纯化和表征 用于鸟苷酸环化酶的激素调节及其功能 重组 这些研究预计将增加我们的 了解环GMP作为第二个基本的作用 信使介导激素的作用，因此，应该提供 改变生物化学和生理反应的机制， 正常和病理状态。

The specific aims of this proposal are to determine the mechanisms of hormone, autocoid and drug regulation of the isoenzyme forms of guanylate cyclase and cyclic GMP synthesis. The precise mechanisms involved in hormone receptor coupling to the different guanylate cyclase isoenzymes will be examined in considerable detail. Experiments will utilize intacts cell cultures and tissues as well as crude and purified components and their reconstitution in cell-free preparations. The effects of atrial natriuretic factor and E. coli heat-stable enterotoxin on cyclic GMP synthesis as well as other ligands and hormones will serve as models and prototypes for these studies. The effects of ATP and other nucleotides and a recently purified protein activator of guanylate cyclase will also be examined. The role of these materials as well as protein phosphorylation and dephosphorylation in guanylate cyclase regulation and hormone coupling to cyclic GMP synthesis will be examined. Methods will include the purification and characterization of coupling factors for hormonal regulation of guanylate cyclase and their functional reconstitution. These studies are expected to increase our understanding of the role of cyclic GMP as a fundamental second messenger to mediate hormone effects and, thus, should provide mechanisms to modify biochemical and physiological responses in normal and pathological states.