BIOSYNTHESIS OF PROLYL HYDROXYLASE

脯氨酰羟化酶的生物合成

• 批准号: 3156112
• 负责人: RICHARD A BERG
• 金额: $ 18.41万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-04-01 至 1988-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3156112
• 关键词: Ehlers Danlos syndrome; atherosclerosis; collagen disorder; fibroblasts; free radicals; gel electrophoresis; genetic transcription; ion exchange chromatography; lysosomes; orphan disease /drug; osteogenesis imperfecta; pulmonary fibrosis /granuloma; rheumatoid arthritis; tissue /cell culture

This study aims at describing the role of lysosomes in regulating collagen production in fibroblasts. The regulation of collagen has clear health-related implications because of the number of human diseases that are associated with a change in collagen metabolism. Acquired disorders such as rheumatoid arthritis, pulmonary fibrosis, cirrhosis, and atherosclerosis, involve significant changes in collagen content and type in affected tissues. The genetic disorders such as the Ehlers Danlos syndromes and Osteogenesis Imperfecta demonstrate various aspects of deficiencies in collagen regulation. Based on recent data from several laboratories that intracellular degradation of newly synthesized collagen is widespread in fibroblasts and that it can in part be inhibited by lysosomal inhibitors, a role for lysosomes in collagen biosynthesis has been implicated. One role of lysosomes has been shown to involve the degradation, intracellularly, of a portion of newly synthesized, structurally defective, collagen. A second role for lysosomes to be tested here is their hypothesized involvement in the translational regulation of collagen synthesis and its secretion. The role of lysosomes is, however, complicated by the presence of at least one non-lysosomal pathway for the basal levels of intracellular degradation of newly synthesized collagen. The first aim of the present proposal is to use cultured chick tendon fibroblasts to determine if procollagen or procollagen fragments are found in lysosomes. The second aim is to examine cultured fibroblasts which are inhibited in theri ability to secrete procollagen to determine if decreased secretion also results in decreased translation, and if this decreased translation is a result of regulation of procollagen production involving lysosomes. The third aim is to determine whether lysosomes may be involved in the uptake of either procollagen or procollagen extension propeptides either alone or by a receptor-mediated process. In order to characterize further intracellular degradation in cultured fibroblasts, the fourth aim is to examine the basal level of degradation to determine if oxygen-derived free radicals and/or neutral proteases are responsible for this important non-lysosomal pathway.

本研究旨在描述溶酶体在调节胶原中的作用 在成纤维细胞中产生。 胶原蛋白的调节有明确的 与健康有关的影响，因为人类疾病的数量， 与胶原代谢的变化有关。 获得性疾病 如类风湿性关节炎、肺纤维化、肝硬化， 动脉粥样硬化，涉及胶原蛋白含量和类型的显着变化 在受影响的组织中。 遗传性疾病，如埃勒斯·丹洛斯 综合征和成骨不全的各个方面， 缺乏胶原蛋白调节。 根据几个国家最近的数据， 新合成的胶原蛋白的细胞内降解 广泛存在于成纤维细胞中，并且可以通过 溶酶体抑制剂，溶酶体在胶原蛋白生物合成中的作用， 被牵连。 溶酶体的一个作用已被证明涉及 降解，细胞内，一部分新合成的， 结构缺陷的胶原蛋白 溶酶体的第二个作用有待检验 这是他们假设参与的翻译调控， 胶原蛋白的合成和分泌。 然而，溶酶体的作用是， 复杂的是存在至少一个非溶酶体途径， 新合成的胶原蛋白的细胞内降解的基础水平。 本建议的第一个目的是使用培养的鸡腱 成纤维细胞，以确定是否发现前胶原或前胶原片段 在溶酶体中。 第二个目的是检查培养的成纤维细胞， 抑制其分泌前胶原的能力，以确定是否降低 分泌也会导致翻译减少，如果翻译减少， 翻译是调节前胶原产生的结果， 溶酶体 第三个目的是确定溶酶体是否可能参与 在前胶原或前胶原延伸前肽的摄取中 单独或通过受体介导的过程。 为了表征 在培养的成纤维细胞中进一步细胞内降解，第四个目的 是检查降解的基础水平，以确定是否氧衍生 自由基和/或中性蛋白酶负责这一重要的 非溶酶体途径