Small Molecule Modulators of the Actin Cytoskeleton

肌动蛋白细胞骨架的小分子调节剂

• 批准号: 6707157
• 负责人: Robert SCOTT LOKEY
• 金额: $ 29.68万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-29 至 2008-09-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6707157
• 关键词: Porifera; Xenopus oocyte; actins; affinity chromatography; biological products; biological signal transduction; combinatorial chemistry; cyclic peptides; cytoskeleton; gel electrophoresis; high throughput technology; intermolecular interaction; molecular assembly /self assembly; molecular dynamics; peptide chemical synthesis; peptide library; phenotype; phorbols; phosphatidylinositols; phosphoproteins; proteomics; western blottings

DESCRIPTION (provided by applicant): Actin filament dynamics in the cell underlie diverse processes, such as neuronal axon guidance, phagocytosis, platelet activation, organelle movement, cell motility, and cancer metastasis. Understanding actin dynamics at the molecular level will give us insight into these processes and may lead to the identification of new drug targets for the inhibition of tumor cell invasion and metastasis. Small molecules in particular have been instrumental in probing fast process such as cytoskeletal dynamics, although most of these have been discovered by serendipity. This research program centers on the systematic discovery of small molecule tools with which to study actin dynamics in mammalian cells. Five aims are proposed: 1) To identify the molecular target of G72*, a cyclic peptide that inhibits phosphoinositol(4,5) bisphosphate (PIP2)-induced actin assembly in Xenopus egg extracts. 2) To generate libraries of cell-permeable cyclic and near, end-capped peptides using diversity-oriented synthetic methods. 3) To purify marine sponge extracts and format the resulting semi-pure fractions for high-throughput screening. 4) To screen synthetic and natural product libraries for compounds that inhibit phorbol-12-myristate-13-acetate (PMA)-induced membrane ruffling in mammalian cells. 5) To identify phosphoproteins and proteins that associate specifically with F-actin assembled during membrane ruffling using a combination of affinity chromatography and proteomic profiling methods.

描述(申请人提供)：肌动蛋白细丝动力学在细胞中的不同过程，如神经元轴突引导、吞噬、血小板激活、细胞器运动、细胞运动和癌症转移。在分子水平上了解肌动蛋白的动力学将使我们深入了解这些过程，并可能导致识别新的药物靶点来抑制肿瘤细胞的侵袭和转移。特别是小分子在探索细胞骨架动力学等快速过程中发挥了重要作用，尽管其中大多数都是偶然发现的。这项研究计划的核心是系统地发现小分子工具，用来研究哺乳动物细胞中的肌动蛋白动力学。我们提出了五个目标：1)确定环肽G72*的分子靶点，G72*是一种环肽，能抑制磷酸肌醇(4，5)二磷酸(PIP2)诱导的非洲爪哇卵提取液中肌动蛋白的组装。2)利用以多样性为导向的合成方法构建细胞渗透性环状多肽和接近末端封端的多肽文库。3)提纯海绵提取物，形成半纯组分，用于高通量筛选。4)筛选合成和天然产物文库，寻找抑制佛波醇-12-肉豆蔻酸酯-13-乙酸酯(PMA)诱导的哺乳动物细胞膜褶皱的化合物。5)结合亲和层析和蛋白质组学方法鉴定膜褶皱过程中形成的磷蛋白和与F-肌动蛋白特异结合的蛋白。