STUDIES OF PROTEINS IN SOLUTION, INTERFACES AND SOLIDS

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，那些不接触溶剂的。包装 在三维拼图的意义上的侧链是 认为在链条的微调中发挥主要作用 构象。包装效果对包装极为敏感。 密度，从而影响蛋白质内的空穴分布。的关注点 这方面的工作在此赠款期间将获得一个 残留物形状的有效测量和确立“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 立即参与生物功能的区域。我们的意图是 尝试开发一种通用的化学标签程序来识别 蛋白质在个体水平上的溶剂可及表面 残留物。蛋白质折叠过程中表面的变化 齐聚反应，或在配体结合过程中反映了大多数 正在研究的分子生物学的生物学功能。因为至少50% 在这个表面上，通常饱和的碳氢化合物是最活跃的 化学物质被要求给它贴上标签。我们会试一试亚甲基。 用于识别标记区域的分析技术将是质量 光谱学。对标记的序列进行排序的最合适的技术 将建立样本。 这些程序将在已知结构的蛋白质上进行验证 核糖核酸酶-S和硫代还蛋白。然后，该方法将应用于 几种细菌的跨膜螺旋的结合和结构 系统，并得到了 与D.M.Engelman、T.A.Steitz和N. 格林德利。后两个问题不是X射线或核磁共振所能解决的 在这个时候的程序。

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.