ASSEMBLY OF APOB48 CONTAINING LIPOPROTEINS

含有脂蛋白的 APOB48 的组装

基本信息

批准号：
2146181
负责人：
M Mahmood Hussain
金额：
$ 10.15万
依托单位：
ALLEGHENY UNIVERSITY OF HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-01-01 至 1999-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2146181
关键词：
animal genetic material tag apolipoproteins blood lipoprotein biosynthesis chylomicrons dietary lipid eating human genetic material tag lipid biosynthesis molecular biology neoplastic cell culture for noncancer research nutrition related tag protein degradation secretion tissue /cell culture transfection transfection /expression vector triglycerides western blottings

项目摘要

The long term objective is to understand the molecular mechanisms involved in the biosynthesis of apoB48-containing lipoproteins. Our working model is that in post-prandial state apoB48-containing lipoproteins are synthesized first as HDL-like primordial lipoproteins and at a later stage the core of these particles is increased to accommodate large amounts of neutral lipids. We hypothesize that the coordinate induction of triglyceride and apoAIV synthesis results in the assembly and secretion of larger chylomicrons. To examine this hypothesis, we will: 1. Develop cell culture that can synthesize large quantities of apoB48- containing lipoproteins and study the effect of increased triglyceride synthesis on the assembly and secretion of these particles. 2. a) Analyze the role of apoAIV in the stabilization of the surface of intracellular nascent particles by expressing apoAIV in Caco-2 cells, cotransfected or not with apoB48, and studying the increase in size of the secreted lipoproteins and b) study the coordinate effects of increased neutral lipid and apoAIV synthesis on lipoprotein assembly and secretion. 3. Determine the rates of intracellular degradation and secretion of apoB48 by pulse-chase analysis of transfected cell lines, perform control experiments to eliminate the possibilities of extracellular degradation, uptake and sequestration of nascent particles and examine the effect of neutral lipid synthesis in conjunction with the overexpression of apoAIV on the rates of intracellular degradation and secretion of apoB48. These studies will provide information about the role of apoAIV in the stabilization of nascent particles and the effect of increased neutral lipid and apoAIV synthesis on intracellular degradation of apoB48. Such information will serve as the groundwork for in vivo studies to modulate lipoprotein secretion in the context of disorders such as hypertriglyceridemia, hypercholesterolemia, and atherosclerosis. The cell lines developed will be valuable in the study of intracellular mechanisms of lipoprotein assembly.

长期目标是了解分子机制参与含ApoB48的脂蛋白的生物合成。我们的工作模型是在野前状态中含有APOB48 脂蛋白首先合成为HDL样原始脂蛋白在后期，这些颗粒的核心增加到容纳大量中性脂质。我们假设协调甘油三酸酯和Apoaiv合成的诱导导致较大的乳糜微粒的组装和分泌。检查一下假设，我们将： 1。发展可以合成大量APOB48-的细胞培养含有脂蛋白并研究甘油三酸酯增加的影响这些颗粒的组装和分泌的合成。 2。a）分析apoaiv在稳定表面的作用细胞内新生颗粒通过在CACO-2细胞中表达apoaiv，与APOB48共转染或不转染，研究大小的增加分泌的脂蛋白和b）研究脂蛋白组装上的中性脂质和载脂合成增加，并增加分泌。 3。确定细胞内降解和分泌的速率通过转染的细胞系的脉冲追踪分析APOB48，执行对照消除细胞外降解的可能性的实验，吸收和隔离新生颗粒并检查与apoaiv的过表达结合的中性脂质合成关于APOB48的细胞内降解和分泌的速率。这些研究将提供有关apoaiv在新生颗粒的稳定和中性增加的影响 ApoB48细胞内降解的脂质和apoAIV合成。这样的信息将作为体内研究调节的基础在疾病中的脂蛋白分泌高甘油三酯血症，高胆固醇血症和动脉粥样硬化。这开发的细胞系对于细胞内研究很有价值脂蛋白组装的机理。