REGULATION OF CYCLIC NUCLEOTIDE FORMATION AND ACTION

这项提案的具体目标是确定 血管紧张素转换酶的激素、自激素和药物调节机制 鸟苷环化酶同工酶和环状GMP合成。 激素受体偶联作用的确切机制 不同的鸟苷环化酶同工酶将在 相当多的细节。实验将利用血管内皮细胞培养 和组织以及粗制和提纯的成分及其 在无细胞制剂中重建。心房肌的作用 利钠因子与大肠杆菌耐热肠毒素对环磷酰胺的影响 GMP合成以及其他配体和激素将作为 这些研究的模型和原型。三磷酸腺苷和丙二醛的作用 其他核苷酸和最近纯化的蛋白激活剂 鸟苷环化酶也将被检测。这些因素的作用 材料以及蛋白质的磷酸化和 鸟苷环化酶调节中的去磷酸化与激素 我们将研究与环状GMP合成的偶联作用。方法将 包括偶联因子的提纯和表征 鸟苷环化酶的激素调节及其功能 重建。这些研究预计将增加我们的 对环状GMP作为第二基础的作用的理解 信使调节荷尔蒙效应，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.