Functional Characterization-Lipoprotein Lipase Mutations

功能表征-脂蛋白脂肪酶突变

• 批准号: 6698838
• 负责人: M. Ilyas Kamboh
• 金额: $ 34.54万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-24 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6698838
• 关键词: atherosclerosis; binding sites; blood lipoprotein metabolism; cardiovascular disorder risk; clinical research; enzyme activity; gel mobility shift assay; gene mutation; genetic enhancer element; genetic polymorphism; genetic regulation; genetic regulatory element; genetic susceptibility; human genetic material tag; human subject; insulin; isozymes; lipoprotein lipase; tissue /cell culture; transcription factor; transfection

DESCRIPTION (provided by applicant): Lipoprotein lipase (LPL) enzyme plays a pivitol role in lipid metabolism by hydrolyzing triglyceride (TG) rich lipoprotein particles. Many diseases, including coronary heart disease (CHD), atherosclerosis and obesity seem to be directly or indirectly related to abnormalities in LPL function. A Hind lll polymorphism in intron 8 of the LPL gene has been shown to be associated with plasma TG and HDL-cholesterol levels and with the risk of CHD. However, the biological mechanism behind this association is unknown. Previous studies have suggested that the intron 8 sequence, which encompasses the Hind lll site as well as intron 9 and non-coding exon 10 of the LPL gene, contain regulatory elements important for LPL transcription and translation. In this proposal we will determine if the nucleotide change in intron 8 by itself is responsible for the effect by disrupting an intronic enhancer element (Aim 1) and what is the role of the 3' region (intron 8, intron 9, exon 10) in regulating LPL transcription (Aim 2). In addition, we have identified a putative insulin response element (IRE) in non-coding exon 10 of the human LPL gene following sequence comparison of LPL genes from various species. The putative IRE sequence was targeted for mutation detection and we identified a 5 bp deletion mutation. We propose to characterize this new mutation further by performing functional and molecular studies (Aim 3). The proposed molecular and functional studies will characterize the functional significance of the LPL Hind lll and exon 10 mutations and may lead to identification and characterization of LPL regulatory elements in intragenic and 3 untranslated region of the gene.

描述（由申请人提供）：通过水解甘油三酸酯（TG）富脂蛋白颗粒，脂蛋白脂肪酶（LPL）酶在脂质代谢中起比维醇的作用。 许多疾病，包括冠心病（CHD），动脉粥样硬化和肥胖似乎与LPL功能异常直接或间接有关。 LPL基因内含子8中的后LLL多态性已​​显示与血浆TG和HDL-胆固醇水平以及CHD风险有关。 但是，这种关联背后的生物学机制尚不清楚。 先前的研究表明，内含子8序列涵盖了后LLL位点以及LPL基因的内含子9和非编码外显子10，其中包含对LPL转录和翻译重要的调节元件。 在此提案中，我们将通过破坏内含子增强子元件（AIM 1）来确定内含子8中的核苷酸变化是否是造成效果的原因，以及3'区域（内含子8，内含子9，外显子10）在调节LPL转录中的作用是什么（AIM 2）。 此外，在对来自各种物种的LPL基因的序列比较后，我们已经确定了人LPL基因的非编码外显子10中的假定胰岛素反应元件（IRE）。 推定的IRE序列是针对突变检测的，我们确定了5 bp的缺失突变。 我们建议通过进行功能和分子研究进一步表征这种新的突变（AIM 3）。 提出的分子和功能研究将表征LPL Hind LLL和外显子10突变的功能意义，并可能导致基因内基因和3个未翻译区域的LPL调节元件的鉴定和表征。