Cell Mutants Defective in Cholesterol Ester Formation

胆固醇酯形成有缺陷的细胞突变体

• 批准号: 7240584
• 负责人: Ta Yuan CHANG
• 金额: $ 42.23万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7240584
• 关键词: Acyl Coenzyme A; Address; Affect; Atherosclerosis; Binding; Cells; Cholesterol; Cholesterol Esters; Cholesterol Homeostasis; Complex; Elements; Endoplasmic Reticulum; Enzymes; Esterification; Event; Fatty Acids; Foam Cells; Glycosphingolipids; Goals; Grant; Human; In Vitro; Knowledge; Latex Bead; Ligands; Lipoproteins; Mammalian Cell; Membrane Biology; Play; Process; Proteins; Research Personnel; Role; Somatic Cell; Sterol O-Acyltransferase; Sterols; Testing; caveolin 1; cholesterol trafficking; crosslink; design; late endosome; macrophage; mutant; programs; research study; response; trafficking

DESCRIPTION (provided by applicant): The tong-term goal of this project is to utilize somatic cell mutants in cholesterol metabolism and macrophage cells to delineate various intracellular cholesterol trafficking events that are relevant to cellular cholesteryl ester accumulation and macrophage foam cell formation in human atherosclerosis. For the proposed granting period, our goals are: (1) To examine the elements involved in endosomal cholesterol trafficking in mammalian cells. (2) To investigate the mechanism by which aggregated lipoproteins cause the resident ER enzyme acyl-coenzyme A: cholesterol acyltransferase 1 (ACAT1) to become more accessible to cholesterol present in various cholesterol-rich compartments. The specific aims are: 1. To identify the elements involved in late endosomal cholesterol trafficking. We will design experiments to address the following questions: a. Does endoCHOL play important roles in maintaining the functions of late endosome in a manner not replaceable by LDL-derived cholesterol? b. Does intracellular caveolin 1 play important roles in late endosomal cholesterol trafficking? 2. To demonstrate NPC1 as a sterol sensing protein in late endosomes. We will design experiments to address the following questions: a. Is the intact cell cross-linking between NPC1 and azocholestanol affected by the presence of NPC2 and/or caveolin 1? b. Does azocholestanol bind to the NPC1 within the sterol sensing domain of NPC1? c. Do NPC1, NPC2, and/or caveolin1 form a protein complex in vitro? Is the in vitro cross-linking between NPC1 and the azocholestanol affected by the presence of other ligands such as fatty acids or glycosphingolipids? 3. To test the possibility that an increase in ER fragmentation/translocation is an early response to certain atherogenic lipoproteins in macrophages. We will design experiments to address the following question: Do aggregated LDLs and latex beads share the same ability to stimulate the endoplasmic reticulum (ER) fragmentation and translocation process in macrophage cells?

描述（由申请人提供）：该项目的倾斜度目标是利用胆固醇代谢和巨噬细胞中的体细胞突变体来描述与细胞内胆固醇酯积累和巨噬细胞泡沫细胞形成的各种细胞内胆固醇运输事件。在拟议的授予期内，我们的目标是：（1）检查哺乳动物细胞中内体胆固醇贩运所涉及的要素。 （2）研究聚集的脂蛋白引起常驻ER酶酰基辅酶A：胆固醇酰基转移酶1（ACAT1）的机制，可以更易于在富含胆固醇的室内存在的胆固醇。具体目的是：1。确定内体胆固醇晚期涉及的要素。我们将设计实验以解决以下问题：内色醇在无法用LDL衍生的胆固醇替代的方式维持后期内体的功能方面起重要作用？ b。细胞内小窝蛋白1是否在晚期内体胆固醇贩运中起重要作用？ 2。在晚期内体中证明NPC1为固醇感应蛋白。我们将设计实验以解决以下问题： NPC1和Azocholestanol之间的完整细胞是否受NPC2和/或Caveolin 1的影响？ b。 NPC1的固醇传感结构域内的硫烷醇是否与NPC1结合？ c。 NPC1，NPC2和/或Caveolin1是否在体外形成蛋白质复合物？ NPC1与偶氮植物之间的体外交联是否受其他配体（例如脂肪酸或糖磷脂）的影响？ 3。为了测试ER碎裂/易位增加是对巨噬细胞中某些动脉粥样硬化脂蛋白的早期反应的可能性。我们将设计实验来解决以下问题：聚集的LDL和乳胶珠是否具有刺激巨噬细胞中内质网（ER）碎片（ER）碎片的能力相同的能力？