Acquisition of an Isothermal Titration Calorimetry System

获得等温滴定量热系统

• 批准号: 0079966
• 负责人: Edward Ferroni
• 金额: $ 5.25万
• 依托单位: Benedictine University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0079966&HistoricalAwards=false
• 关键词: Acquisition; Isothermal; Titration; Calorimetry; System

An isothermal titration calorimetry (ITC) system from MicroCal Incorporated will be used at Benedictine University for biochemical enzymatic studies. The system includes a titration calorimeter and computer for automatic control of the instrument. When two substances interact or combine (noncovalently or covalently), the absorption or evolution of heat occurs. The MicroCal ITC system offers a sensitive and rapid way to determine the amount of heat released or taken up by such a reaction. Computer analysis of the data acquired yields the desired binding or interaction constants. Initially the system will be used in three different projects. Project I involves thestudy of the binding of dihydroxyacetone phosphate (DHAP) analogues to rabbit muscle aldolase (RAMA). This enzyme, which is inexpensive and readily available commercially, catalyzes the addition of DHAP to a variety of aldehydes. As a result, RAMA has been used for the organic synthesis of a number of novel compounds. The analogues will be synthesized and the ability of these compounds to bind to aldolase as a function of their structure will be determined using the ITC system. The information should shed further light on the binding specificity of the aldolase active site and provide further details concerning the enzyme mechanism. Project II will investigate the ability of synthetic crown ethers to bind cesium ions. Cesium-137 is a radioactive byproduct of nuclear reactors. Removal of the cesium from wastewater can be done by extraction with inclusion compounds (e.g., crown ethers). The isothermal titration calorimetry system will be used to rapidly screen the synthetic crown ethers for their ability to bind the cesium ion. The binding of peptides to fibrinogen or the fibrinogen receptors of platelets will be investigated in project III. Peptides will be tested for their ability to bind to fibrinogen or cellular receptors using the isothermal titration calorimetry system. The successful completion of this project will make important contributions to the understanding of the fibrinogen/receptor interaction. Faculty and students will use this instrumentation in interdisciplinary research projects, classroom exercises

本笃会大学将使用MicroCal公司的等温滴定量热法（ITC）系统进行生化酶研究。该系统包括一个滴定量热仪和计算机自动控制的仪器。当两种物质相互作用或联合收割机（非共价或共价）时，就会发生吸热或放热。MicroCal ITC系统提供了一种灵敏、快速的方法来确定这种反应释放或吸收的热量。计算机分析所获得的数据产生所需的结合或相互作用常数。 该系统最初将用于三个不同的项目。第一个项目是研究磷酸二羟丙酮（DHAP）类似物与兔肌肉醛缩酶（拉马）的结合。这种酶价格便宜，容易在商业上获得，催化DHAP加成到各种醛中。因此，拉马已被用于许多新化合物的有机合成。将合成类似物，并将使用ITC系统确定这些化合物与醛缩酶结合的能力（作为其结构的函数）。该信息应进一步阐明醛缩酶活性位点的结合特异性，并提供有关酶机制的进一步细节。项目二将研究合成冠醚结合铯离子的能力。铯-137是核反应堆的放射性副产品。从废水中除去铯可以通过用包合化合物（例如，冠醚）。等温滴定量热系统将用于快速筛选合成冠醚的能力，结合铯离子。将在项目III中研究肽与纤维蛋白原或血小板的纤维蛋白原受体的结合。将使用等温滴定量热法系统检测肽与纤维蛋白原或细胞受体结合的能力。该项目的成功完成将对纤维蛋白原/受体相互作用的理解做出重要贡献。教师和学生将在跨学科的研究项目，课堂练习中使用这种仪器