PITUITARY KININOGENASES--PROPERTIES AND REGULATION

垂体激肽酶——特性和调节

• 批准号: 3447244
• 负责人: CLAUD A POWERS
• 金额: $ 5.61万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3447244
• 关键词: adrenalectomy; adrenocorticotropic hormone; affinity chromatography; cell components; density gradient ultracentrifugation; desoxycorticosterone; enzyme substrate; gel electrophoresis; high performance liquid chromatography; hormone regulation /control mechanism; hypothalamus; kallikreins; male castration; melanocyte stimulating hormone; molecular weight; opiate alkaloid; pituitary gland; radioimmunoassay; species difference; testosterone; thyroidectomy; thyroxine

The objective of the proposed research is to study the biochemical properties, subcellular distribution, and physiological regulation and significance of kiningenerating proteinases (kininogenases) I have recently detected in rat and porcine pituitaries. Rat and porcine pituitary kininogenases will be solubilized, and purified using a combination of affinity and gel-filtration chromatography. Purified kininogenases will be characterized with regard to molecular weight, specificity and kinetics of substrate hydrolysis, sensitivity to synthetic and natural inhibitors, and ability to cleave intact rat pro-opiomelanocortin into its hormonal fragments and generate opioid or melanocyte-stimulating peptides from ACTH or Beta-lipotropin. Rat and porcine pituitaries will be homogenized in sucrose buffer and various subcellular fractions prepared using a combination of differential centrifugation and discontinuous sucrose density gradients; kininogenase activity in the various subcellular fractions will be determined using synthetic and natural substrates for kininogenases. In the rat, responses of pituitary kininogenases to various endocrine and pharmacological manipulations will be examined both in vitro and in vivo in order to characterize the factors which may regulate pituitary kininogenases and their function. In addition, pituitaries of various mouse strains, amphibians, and reptiles will be compared with respect to kininogenase activity. The proposed research should elucidate the characteristics of kininogenases associated with various pituitary lobes in different species and suggest how kininogenases may regulate pituitary function. Such studies should lay the foundation for future research on the role of kininogenses and other proteases in pituitary function using techniques of pharmacology, endocrinology, and molecular biology. The pituitary promises to be a model system for studying how specific proteases regulate the post-tranalational processing of secretory proteins. Such research may ultimately illuminate the involvement of proteases in clinical syndromes characterized by an altered production of peptide hormones or othe protein products.

这项研究的目的是研究 性质、亚细胞分布和生理调节， 产生激肽的蛋白酶（激肽原酶）的意义我最近 在大鼠和猪的垂体中检测到。 大鼠和猪垂体 激肽原酶将被溶解，并使用以下的组合进行纯化： 亲和层析和凝胶过滤层析。 纯化的激肽原酶将 在分子量、特异性和动力学方面表征 底物水解，对合成和天然抑制剂的敏感性，以及 将完整的大鼠阿黑皮素原切割成其激素的能力 片段并从ACTH产生阿片样物质或黑素细胞刺激肽 或β-促脂素。 将大鼠和猪垂体匀浆， 蔗糖缓冲液和使用 差速离心和不连续蔗糖的组合 密度梯度;激肽原酶活性的各种亚细胞 将使用合成和天然底物测定馏分， 激肽原酶 在大鼠中，垂体激肽原酶对各种 内分泌和药理学操作将在体外进行检查， 为了表征可能调节 垂体激肽原酶及其功能。 此外， 各种小鼠品系，两栖动物和爬行动物将与 关于激肽原酶活性。 拟议的研究应阐明 与各种垂体相关的激肽原酶的特性 不同物种的叶，并提出激肽原酶如何调节 垂体功能 这样的研究应该为将来的研究奠定基础。 激肽原酶等蛋白酶在垂体中作用的研究 利用药理学、内分泌学和分子生物学技术， 生物学 脑垂体有望成为研究 特定的蛋白酶调节分泌的翻译后加工， proteins. 这样的研究最终可能会阐明 蛋白酶在临床综合征的特点是改变生产 肽激素或其他蛋白质产品。