STUDIES OF PROTEINS IN SOLUTION, INTERFACES AND SOLIDS

溶液、界面和固体中蛋白质的研究

• 批准号: 6107471
• 负责人: FREDERIC M RICHARDS
• 金额: $ 24.68万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6107471
• 关键词: biomaterial interface interaction; chemical synthesis; mass spectrometry; pancreatic ribonuclease; photolysis; protein folding; protein sequence; solid state; thioredoxin; water solution

The general fold of a polypeptide chain appears to be largely controlled by the interior residues, those that do not contact the solvent. packing of the side chains in the sense of a three dimensional jigsaw puzzle is thought to play a major role in fine adjustmentS of the chain conformation. packing effects are exquisitely sensitive to packing density, and thus to cavity distribution within the protein. The focus of this aspect of the work during this grant period will be to obtain a useful measure of residue shape and establish the importance of "fit" in the folding process. Strangely enough the answer is not obvious. The second focus of the work will be on the surface of the protein, the region immediately involved in biological function. It is our intent to try to develop a general chemical labeling procedure for identifying the solvent accessible surface of a protein at the level of individual residues. The changes in the surface during protein folding, during oligomerization reactions, or during ligand binding reflect most of the biological functions under study in molecular biology. Since at least 50% of this surface is normally saturated hydrocarbons the most reactive chemical species are required to label it at all. We will try methylene. The analytical technique for identifying labeled areas will be mass spectroscopy. The most appropriate technique for sequencing the labeled samples will be established. The procedures will be validated on proteins of known structures starting with ribonuclease-S and thiroedoxin. The method will then be applied to the association and structure of the transmembrane helices in several systems, and to the higher order structures of delta-gamma resolvase in collaboration with the laboratories of D.M. Engelman, T.A. Steitz, and N. Grindley. These latter problems are beyond the reach of X-ray or NMR procedures at this time.

多肽链的一般折叠似乎在很大程度上受到控制 通过内部残留物，那些不接触溶剂的残留物。包装 从三维拼图的意义上讲，侧链的拼图是 被认为在链条的精细调整中发挥了重要作用 构象。包装效果对包装非常敏感 密度，从而在蛋白质内进行腔分布。重点 在此赠款期间的工作这一方面将是获得 残留形状的有用度量，并确定“拟合”的重要性 折叠过程。奇怪的是，答案并不明显。 工作的第二个焦点将是蛋白质表面 立即参与生物功能的区域。这是我们的意图 尝试制定一般的化学标签程序来识别 蛋白质水平的蛋白质的溶剂表面 残留物。蛋白质折叠期间表面的变化，在 寡聚反应，或在配体结合期间反映了大多数 分子生物学研究的生物学功能。由于至少50％ 该表面通常是饱和的碳氢化合物 完全需要化学物质来标记它。我们将尝试亚甲基。 用于识别标记区域的分析技术将是质量 光谱法。测序标记的最合适的技术 将建立样品。 该过程将在已知结构的蛋白质上进行验证 与核糖核酸酶-s和硫氧还蛋白一起使用。然后将该方法应用于 几个跨膜螺旋的关联和结构 系统，以及在Delta-Gamma分解的高阶结构中 与D.M.实验室合作恩格曼（T.A.） Steitz和N. 格林德利。这些后一种问题超出了X射线或NMR的影响 目前的程序。