Regulation of caveolae internalization

小窝内化的调节

• 批准号: 7002236
• 负责人: John K Zehmer
• 金额: $ 4.88万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7002236
• 关键词: RNA interference; SDS polyacrylamide gel electrophoresis; actins; caveolas; caveolins; cell membrane; chromatography; cytoplasm; endocytosis; intracellular transport; ligands; postdoctoral investigator; protein kinase C; protein structure function; receptor binding

DESCRIPTION (provided by applicant): The proposed research will help elucidate the molecular mechanisms and regulatory elements governing endocytosis via caveolae. Specifically, an in vitro caveolae budding assay will be used to determine the roles of actin and caveolin-1 in stabilizing caveolae at the cell surface. Furthermore, the budding assay and column chromatography will be used to identify the cytosolic factors required for caveolae budding. The roles of PKC (alpha) and the serum deprivation response protein in regulating caveolae budding will be investigated using RNAi knockdown and the in vitro assay. Furthermore, the influence of ligand binding on receptor internalization by caveolae will be investigated. All factors identified using the in vitro assay will be tested for their relevance in whole cells. Many disease-causing agents including bacteria (e.g. Chlamidia trechomatis) viruses (e.g. the Ebola and Marburg filoviruses), and bacterial toxins (e.g. cholera toxin) are internalized through caveolae, thus bypassing pathways that lead to degradation in lysosomes. Therefore, a deeper understanding of the regulation of caveolae budding could lead to clinical applications.

描述（由申请人提供）： 拟议的研究将有助于阐明通过小窝控制内吞作用的分子机制和调控元件。具体来说，体外小窝出芽测定将用于确定肌动蛋白和小窝蛋白-1 在稳定细胞表面小窝中的作用。此外，出芽测定和柱色谱将用于鉴定小窝出芽所需的胞质因子。将使用 RNAi 敲低和体外测定来研究 PKC (α) 和血清剥夺反应蛋白在调节小窝出芽中的作用。此外，还将研究配体结合对小窝受体内化的影响。使用体外测定确定的所有因素都将测试其在全细胞中的相关性。许多致病因子，包括细菌（例如沙眼衣原体）、病毒（例如埃博拉病毒和马尔堡丝状病毒）和细菌毒素（例如霍乱毒素）通过小窝内化，从而绕过导致溶酶体降解的途径。因此，对小窝出芽调节的更深入了解可能会导致临床应用。