LIPOPEPTIDE STRUCTURE AND FUNCTION

脂肽结构和功能

• 批准号: 3305965
• 负责人: JEFFREY M. BECKER
• 金额: $ 20.86万
• 依托单位: UNIVERSITY OF TENNESSEE KNOXVILLE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3305965
• 关键词: Saccharomyces cerevisiae; endocytosis; endopeptidases; enkephalins; farnesyl compound; fungal genetics; genetic library; genetic strain; hormone inhibitor; hormone receptor; hydrolase; molecular cloning; nonblood lipoprotein; nucleic acid sequence; peptide chemical synthesis; peptide hormone; peptide hormone analog; phenotype; pheromone; polymerase chain reaction; posttranslational modifications; protein degradation; protein purification; protein structure function; receptor binding; site directed mutagenesis; stimulant /agonist

Hormones play a vital role in the regulation of cellular activity. An understanding of the initial event of hormone action, the interaction of hormone with its receptor, is necessary for designing antagonists and superactive agonists useful in therapy of endocrinological disorders. We propose to study the pheromone a-factor (YIIKGVFWDPAC[FARNESYL]-OCH3) as a model for lipopeptide structure and function. Farnesylation of proteins is a newly-discovered, post-translational modification that has been shown to play a critical role in directing modified peptides and proteins to the plasma membrane. Examples of eukaryotic proteins that contain the post-translational polyisoprenoid modification include nuclear lamins, trimeric G proteins, and the RAS family of oncoproteins. Thus, the study of the interaction of a-factor with its receptor and target cell should provide novel information to elucidate the molecular events involved between lipopeptides and cells. We propose to use a combined approach of chemical synthesis and oligonucleotide-directed, random mutagenesis to discover antagonists and superactive agonists. By developing an assay to study the interaction of a-factor and a-factor analogs with its receptor we hope to delineate those residues and molecular groupings involved in pheromone binding. By studying and cloning a newly discovered a-factor protease we hope to uncover the role of lipopeptide degradation in desensitization and recovery of the target cell from pheromone action. By determining whether and how a-factor is internalized we hope to elucidate the cellular fate of hydrophobic molecules subsequent to cellular binding. We believe that the system we propose to investigate will be useful for drug screening for the discovery of important new pharmacological reagents to treat a variety of human diseases.

激素在调节细胞活性中起着至关重要的作用。 对激素作用的初始事件的理解，相互作用 具有其受体的激素，对于设计拮抗剂和 超级活跃的激动剂可用于治疗内分泌疾病。 我们 建议研究信息素A因子（yiikgvfwdpac [farnesyl] -och3） 脂肪肽结构和功能的模型。 法尼的化 蛋白质是一种新发现的翻译后修饰，具有 被证明在指导改良肽和 蛋白质到质膜。 真核蛋白的例子 包含翻译后的多分球菌修饰包括 核层粘蛋白，三聚体G蛋白和癌蛋白的RAS家族。 因此，研究A因子与其受体的相互作用和 靶细胞应提供新的信息以阐明分子 脂肽和细胞之间涉及的事件。 我们建议使用 化学合成和寡核苷酸指导的联合方法 随机诱变，发现拮抗剂和超活动激动剂。 经过 开发一个研究A因子和A因子的相互作用的测定法 与其受体的类似物，我们希望描述这些残留物和 分子分组参与信息素结合。 通过学习和 克隆一个新发现的A因子蛋白酶，我们希望能揭开该角色 靶向和恢复脂肪肽降解的降解 来自信息素动作的细胞。 通过确定A-Factor是如何以及如何 内在化，我们希望阐明疏水的细胞命运 分子在细胞结合之后。 我们相信我们的系统 建议进行调查将对筛查的药物筛查有用 发现重要的新药理试剂来治疗各种 人类疾病。