Studies on Protein Lipidation

蛋白质脂化研究

• 批准号: 9707092
• 负责人: Howard C Hang
• 金额: $ 7.87万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9707092
• 关键词: Acyltransferase; Address; Alkynes; Animal Model; Antiviral Agents; Applications Grants; Bacteria; Biochemical; Bioinformatics; Biological; Biological Assay; Biology; Cell Line; Cells; Cellular biology; Chemicals; Complex; Cysteine; Dendritic Cells; Detection; Eukaryota; Fatty Acids; Genomics; Goals; Grant; Host Defense; Host resistance; Human; Immunity; In Vitro; Infection; Influenza A virus; Integral Membrane Protein; Integration Host Factors; Interferons; Label; Laboratories; Life; Ligation; Lipids; Mammalian Cell; Membrane; Metabolic; Methods; Mus; Palmitic Acids; Pathway interactions; Phenotype; Protein Analysis; Protein Biochemistry; Protein Isoforms; Protein S; Proteins; Proteomics; Reagent; Reporter; Resistance; Role; Series; Signal Pathway; Site; Virus Diseases; Yeasts; base; biophysical analysis; cell type; combat; esterase; fatty acylation; influenzavirus; microbial; mutant; novel; palmitoylation; predictive tools; reconstitution; tool

PROJECT SUMMARY Lipid chemical reporters and bioorthogonal ligation methods developed by my laboratory have provided new opportunities to investigate the functions of lipid-modified proteins in biology. Our proteomic analysis of fatty- acylated proteins in dendritic cells using fatty acid chemical reporters and bioorthogonal chemical proteomics has revealed a new role for protein S-fatty-acylation in host defense against viral infections. We discovered that S-fatty-acylation of membrane-proximal cysteines on murine IFITM3 is crucial for its membrane localization and antiviral activity against influenza A virus. The mechanisms that control S-fatty-acylation of human IFITM proteins have not been evaluated and will be addressed in this grant proposal. Aim 1 will evaluate the S-fatty-acylation levels, sites and function of human IFITM isoforms in different cell types/states of activation. Aim 2 describes the characterization of fatty acids covalently attached to human IFITM3 in cells. Aim 3 describes site-specific lipidation and reconstitution of IFITM3 in vitro for detailed biochemical and biophysical studies. Determining the mechanisms that control S-fatty-acylation IFITM3 function is crucial for understanding host immunity and may reveal new strategies for combatting virus infection in humans. The chemical approaches described in this grant should provide new reagents and methods for studying fatty- acylated proteins.

项目摘要 本实验室开发的脂质化学报告分子和生物正交连接方法为生物医学领域提供了新的 研究脂质修饰蛋白质在生物学中的功能的机会。我们的蛋白质组学分析- 使用脂肪酸化学报告和生物正交化学蛋白质组学在树突状细胞中的酰化蛋白质 揭示了蛋白S-脂肪酰化在宿主防御病毒感染中的新作用。我们发现 小鼠IFITM 3上膜近端半胱氨酸的S-脂肪酰化对于其膜 定位和抗甲型流感病毒的抗病毒活性。控制S-脂肪酰化的机制 人IFITM蛋白尚未被评估，将在本拨款提案中予以解决。目标1将 评价人IFITM亚型在不同细胞类型/状态下的S-脂肪酰化水平、位点和功能， activation.目的2描述了在细胞中共价连接至人IFITM 3的脂肪酸的表征。 目的3描述了IFITM 3在体外的位点特异性脂化和重建，用于详细的生物化学和生物化学性质。 生物物理学研究确定控制S-脂肪酰化IFITM 3功能的机制对于 了解宿主免疫力，并可能揭示对抗人类病毒感染的新策略。的 这项资助中描述的化学方法应该为研究脂肪酸提供新的试剂和方法， 酰化蛋白质