PHYSICOCHEMICAL STUDIES OF BIOCHEMICAL REACTIONS

生化反应的物理化学研究

• 批准号: 2166058
• 负责人: JUDITH M STURTEVANT
• 金额: $ 16.06万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1974
• 资助国家: 美国
• 起止时间: 1974-10-15 至 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2166058
• 关键词: calorimetry; chemical reaction; enzyme structure; lipid structure; nucleic acid metabolism; nucleic acid structure; protein denaturation; protein isoforms; protein structure; temperature; thermodynamics

The broad aim in this project is the study of biological and biochemical processes using physicochemical techniques. As in the past, our efforts will be centered on the determination, primarily by calorimetric methods and the interpretation of the thermodynamics of such processes. A major fraction of our work will involve study of thermally-induced processes in proteins, nucleic acids and lipids by means of high sensitivity differential scanning calorimetry (DSC). The power and applicability of this technique is at present rapidly expanding. For example, DSC affords the best means for checking the thermal stability of proteins engineered to perform some specific pharmaceutical function, and for acquiring thermodynamic data which may suggest specific ways to improve the thermal stability. Our DSC work on proteins will remain centered on studying the effects of mutations using materials supplied to use by a wide range of collaborators. Included will be, among others, staphylococcal nuclease, streptomyces subtilisin inhibitor (SSI), thioredoxin and cytochrome b562. DSC work on nucleotides will include study of the unfolding of synthetic triple stranded DNA, and of various nucleotide-protein complexes also studies by ITC. DSC gives an excellent means for studying the phase transitions of lipids. Work on pure synthetic lipids of unusual structure will continue. Studies of the effects of added materials (Ca++, Mg++, AlCl3, nitrophenyl esters) on the thermotropic behavior of lipid bilayers will also be continued. We are making constantly increasing use of isothermal situation calorimetry (ITC), in studying the binding of ligands to proteins and nucleic acids, the interactions of proteins with nucleic acids, for example, the trp repressor-trp operator interaction, and other biochemical reactions.

该项目的主要目的是研究生物和生物化学 使用物理化学技术的过程。 与过去一样，我们的努力 将集中在测定，主要是通过量热法 以及对这些过程的热力学解释。 我们工作的一个主要部分将涉及研究热诱导 蛋白质、核酸和脂质中的过程， 灵敏度差示扫描量热法（DSC）。 电力和 这种技术的适用性目前正在迅速扩大。 为 例如，DSC提供了检查热稳定性的最佳方法 蛋白质被改造以执行某些特定的药物功能， 以及用于获取热力学数据，这些数据可以建议具体的方法， 提高热稳定性。 我们在蛋白质方面的DSC工作将继续 集中研究突变的影响， 供广泛的合作者使用。其中包括， 葡萄球菌核酸酶，链球菌枯草杆菌蛋白酶抑制剂（SSI）， 硫氧还蛋白和细胞色素b562。 关于核苷酸的DSC工作将包括研究合成核苷酸的解折叠 三链DNA和各种核苷酸-蛋白质复合物也 ITC的研究。 DSC为研究相变提供了极好的手段， 脂质。 对结构不寻常的纯合成脂质的研究将 继续 研究了添加材料（Ca++，Mg++，AlCl 3， 硝基苯基酯）对脂双层的热致行为的影响将 也继续。 我们正在不断增加使用等温情况 量热法（ITC），在研究配体与蛋白质的结合， 蛋白质与核酸的相互作用， 例如，trp阻遏物-trp操纵子相互作用，以及其他 生化反应。