TRANSFORMING GROWTH FACTOR B1

转化生长因子 B1

• 批准号: 3459131
• 负责人: LARRY E GENTRY
• 金额: $ 9.14万
• 依托单位: UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 1993-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3459131
• 关键词: cell differentiation; cell growth regulation; cysteine; endonuclease; gel electrophoresis; gene deletion mutation; gene mutation; high performance liquid chromatography; hormone binding protein; immunochemistry; immunoprecipitation; laboratory mouse; natural gene amplification; nucleic acid hybridization; point mutation; protein biosynthesis; protein engineering; protein structure function; proteolysis; radionuclides; tissue /cell culture; transforming growth factors

Transforming growth factor-beta-1 (TGF-beta1) belongs to a closely related family of polypeptides with potent cellular modulating activities. This growth factor molecule appears to be intimately associated with cell growth and differentiation, and may play pivotal roles in the autocrine regulation of these processes. Although numerous studies have addressed important cellular and physiological functions of TGF-beta1, essentially no information concerning its structure/function analysis have been reported. Consequently, our current understanding of functional domains and potentially important structural aspects is restricted to classical protein chemistry and cDNA sequence analysis. The overall objective of this proposal is to define and investigate structural features of TGF-beta1 important for biological function. To achieve this goal, three principle DNA mutagenesis schemes, in frame insertion, deletion, and site-directed mutagenesis, will be employed to generate a series of mutant TGF-beta1 polypeptides from the cloned simian pre-pro-TGF- beta1 cDNA. The mutant TGF-beta1 cDNA will be expressed in mammalian tissue culture cells and the altered TGF-beta1 polypeptides extensively analyzed by biochemical and immunochemical techniques. These assays will address the effect of the introduced genetic lesions on synthesis, maturation, and receptor binding activity. Particular emphasis will be placed on proteolytic maturation and on formation of a latent biological complex, properties of TGF-beta1 which may play fundamental roles in the regulation of activity. Mutant polypeptides will also be examined for biological activity in standard growth proliferation and inhibition assays as well as cellular differentiation systems. Finally, the precursor sequences of the TGF-beta1 polypeptide will be expressed independent of the mature growth factor and the biological activity associated with this portion assessed by several in vitro and in vivo bioassays. The above proposed studies should allow for a thorough genetic dissection of the TGF-beta1 polypeptide. In addition to identifying important structural and functional domains, these studies should address several characteristic features of TGF- beta1 including the role of the precursor, individual cysteine residues, and protelytic processing for biological function of TGF- beta1. Moreover, information obtained from these studies may identify regions of the mature growth factor essential for its receptor binding properties.

转化生长因子-β-1 (TGF-β1) 属于 密切相关的多肽家族，具有有效的细胞活性 调节活动。 这种生长因子分子似乎是 与细胞生长和分化密切相关，并且 可能在这些细胞的自分泌调节中发挥关键作用 流程。 尽管许多研究已经解决了重要的 TGF-β1 的细胞和生理功能，基本上没有 有关其结构/功能分析的信息已 报道称。 因此，我们目前对函数式的理解 领域和潜在的重要结构方面受到限制 经典蛋白质化学和 cDNA 序列分析。 这 该提案的总体目标是定义和调查 TGF-β1 的结构特征对生物学很重要 功能。 为了实现这一目标，DNA 诱变的三个原则 方案，帧插入、删除和定点 诱变，将被用来产生一系列突变体 来自克隆的猿猴前原 TGF- 的 TGF-β1 多肽 β1 cDNA。 突变型 TGF-β1 cDNA 将在 哺乳动物组织培养细胞和改变的 TGF-β1 通过生化和分析广泛分析多肽 免疫化学技术。 这些测定将解决影响 引入的基因损伤对合成、成熟和 受体结合活性。 将特别强调 蛋白水解成熟和潜在生物的形成 TGF-β1 的复杂特性可能发挥着基础作用 活动调节中的作用。 突变的多肽也会 在标准生长中检查生物活性 增殖和抑制测定以及细胞 差异化系统。 最后，前体序列 TGF-β1 多肽的表达独立于 成熟生长因子及其相关的生物活性 该部分通过多项体外和体内生物测定进行评估。 这 上述建议的研究应允许进行彻底的遗传研究 TGF-β1 多肽的解剖。 此外 识别重要的结构和功能域，这些 研究应解决 TGF-β 的几个特征 beta1 包括前体、单个半胱氨酸的作用 残基和 TGF-生物功能的蛋白水解加工 测试版1。 此外，从这些研究中获得的信息可能 确定其成熟生长因子所必需的区域 受体结合特性。