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 由内部残留物，即那些不接触溶剂的残留物。包装 三维拼图意义上的侧链是 被认为在链条的微调中发挥着重要作用 构象。包装效果对包装非常敏感 密度，从而影响蛋白质内的空腔分布。重点是 在此赠款期内这方面的工作将是获得 残留物形状的有用测量并确定“适合”的重要性 折叠过程。奇怪的是，答案并不明显。 这项工作的第二个重点将是蛋白质的表面，即 直接参与生物功能的区域。我们的目的是 尝试制定通用化学标签程序来识别 个体水平上蛋白质的溶剂可及表面 残留物。蛋白质折叠过程中表面的变化 寡聚反应，或配体结合过程反映了大部分 分子生物学中研究的生物功能。因为至少有 50% 该表面的通常是饱和烃，反应性最强 根本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 光谱学。对标记的测序最合适的技术 将建立样本。 该程序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。然后该方法将被应用到 几种跨膜螺旋的关联和结构 系统，以及 δ-γ 解离酶的高阶结构 与 D.M. 实验室的合作恩格曼，T.A.施泰茨，N. 格林德利。后面这些问题超出了 X 射线或 NMR 的范围 此时的手续。