SCANNING CALORIMETRIC STUDY OF APOLIPOPROTEIN A AND PANCREATIC LIPASE

载脂蛋白A和胰腺脂肪酶的扫描量热研究

• 批准号: 5211672
• 负责人: Romonia Renee REAMS
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5211672
• 关键词: acidity /alkalinity; animal tissue; apolipoproteins; calorimetry; lipase; protein folding; protein purification; thermodynamics

The immediate goal of this research is to study the thermal unfolding of two biomedically important proteins: apolipoprotein (a) and porcine pancreatic lipase at various pH values, solvent conditions and protein concentrations. Apolipoprotein (a), a cholesterol-ester/LDL-like particle was selected for analysis because it has been found to be highly atherogenic (that is, associated with atherosclerotic cardiovascular disease) and has a fascinating structure composed of 17 almost identical domains. At the carboxy terminus, apolipoprotein (a) has single copies of sequences resembling the protease domain of plasminogen. Apolipoprotein (a) is not commercially available but milligram quantities of it are available to us from a confirmed source. Porcine pancreatic lipase has been selected for analysis because of its function; which is to catalyze the hydrolysis of ester bonds of nonpolar dietary long-chain triglycerides. Students will be trained in protein purification techniques and will (1) purify pancreatic lipase from the pancreas of freshly slaughtered hogs, (2) characterize pancreatic lipase using enzyme assays and biochemical methods and (3) study the nature of the proteins' thermal unfolding using differential scanning calorimetry.

这项研究的直接目的是研究 两种生物医学重要的蛋白质：载脂蛋白（A）和猪 胰腺脂肪酶在各种pH值，溶剂条件和蛋白质上 浓度。 载脂蛋白（A），胆固醇酯/LDL样 选择粒子进行分析，因为已经发现它是高度的 动脉粥样硬化（即与动脉粥样硬化心血管有关 疾病），具有迷人的结构，由17个几乎相同 域。 在羧基终端，载脂蛋白（a）具有单份 类似纤溶酶原蛋白酶结构域的序列。 载脂蛋白（a）不能在市售，但毫克数量 它可以从确认的来源向我们使用。 猪胰腺 由于其功能，已选择脂肪酶进行分析。就是 催化非极性饮食长链的酯键的水解 甘油三酸酯。 学生将接受蛋白质净化的培训 技术和意志（1）从胰腺纯化胰腺脂肪酶 新鲜屠宰的猪，（2）使用酶来表征胰腺脂肪酶 测定和生化方法，（3）研究蛋白质的性质 使用差分扫描量热法展开热量。