INFLUENZA HEMAGGLUTININ AS A MODEL ACYLPROTEIN

作为酰基蛋白模型的流感血凝素

• 批准号: 3455027
• 负责人: CLAYTON W. NAEVE
• 金额: $ 9.4万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-01-01 至 1994-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3455027
• 关键词: acylation; cell membrane; chemical models; chloramphenicol acetyltransferase; covalent bond; exocytosis; fatty acid transport; fatty acids; fatty acylation; genetic translation; hemagglutination test; hydroxylamine; influenza; membrane fusion; membrane permeability; membrane proteins; microorganism hemagglutinin; molecular cloning; mutant; site directed mutagenesis; synthetic peptide; transport proteins; virus protein

The covalent attachment of fatty acid moieties to proteins is a widespread postranslational modification of many viral and cell proteins. Fatty acids have been suggested to be important in receptor assembly, protease protection, growth regulation, morphogenesis, membrane anchorage and membrane fusion. However, the most fundamental aspects of acylation and the role fatty acids play in protein structure and function are not yet understood. One of the best characterized membrane glycoproteins is also an acylprotein; the influenza virus hemagglutinin (HA). Recent studies have implicated fatty acid on the HA in the process of membrane fusion, an event crucial for virus infectivity and a process occurring continuously in eukaryotic cells. I propose to use site-directed mutagenesis of a cloned HA gene and cell-free acyltransferase assays using synthetic peptide substrates to identify fundamental structural requirements for protein fatty acid acylation and to characterize the functional role of fatty acid in membrane fusion and budding. These experiments are designed to test the hypothesis that fatty acid on the HA can significantly influence structural or biological properties of the molecule. The results obtained will increase our understanding of this important human pathogen and also improve our view of a widespread post-translational modification beyond the virological context to many cell proteins.

脂肪酸部分与蛋白质的共价连接是一种 许多病毒和细胞的广泛的翻译后修饰 proteins. 脂肪酸被认为是重要的， 受体组装，蛋白酶保护，生长调节， 形态发生、膜锚定和膜融合。 然而，在这方面， 酰化的最基本方面和脂肪酸的作用 氨基酸在蛋白质结构和功能中的作用还不清楚， 明白 最具特征的膜糖蛋白之一 也是一种酰基蛋白;流感病毒血凝素（HA）。 最近的研究表明，脂肪酸对HA的影响， 膜融合过程，这是病毒 传染性和一个过程不断发生在真核生物 细胞 我建议使用克隆HA的定点突变 使用合成肽的基因和无细胞酰基转移酶测定 基板，以确定基本的结构要求， 蛋白质脂肪酸酰化和表征功能 脂肪酸在膜融合和出芽中的作用。 这些 设计实验来检验脂肪酸 对HA的结构或生物学影响 分子的性质。 所取得的成果将增加 我们对这种重要的人类病原体的了解， 改进了我们对广泛翻译后修饰的看法 超越病毒学的范畴，应用于许多细胞蛋白。