ENZYME REACTION INTERMEDIATES--A NEW APPROACH

化合物的立体化学和电子结构的测定 酶反应中间体中的底物和活性中心残基 提出了用冷冻液稳定化的方法。反应中间体 在零下温度下稳定的有机-水共溶剂混合物 将为EPR和Endor方法的光谱研究做准备。这个 顺磁位与质子及其他核素的径向分离 在顺磁位置附近的位置将根据 底物的EPR和Endor吸收以及催化必需的 活性中心残留物。然后，这些结果将被用于分配 结合在底物中的准确的催化生成构型 计算机控制分子在酶活性部位的应用 图形技术。这些方法旨在确定局部结构 动力学活性反应中的酶活性部位和底物 用于评估酶的结构和电子基础的中间体 在详细的分子水平上发挥作用。

The determination of the stereochemistry and electronic structure of substrates and active site residues in reaction intermediates of enzymes stabilized by cryosolvent methods is proposed. Reaction intermediates stabilized at subzero temperatures in organic-aqueous cosolvent mixtures will be prepared for spectroscopic studies by EPR and ENDOR methods. The radial separation between paramagnetic sites and protons and other nuclides in the vicinity of the paramagnetic sites will be determined on the basis of the EPR and ENDOR absorptions of substrates and catalytically essential active site residues. These results will then be employed to assign the exact catalytically productive configurations of substrates bound in the active site of enzymes through application of computer controlled molecular graphics techniques. The methods are designed to determine local structure of enzymic active sites and of substrates in kinetically competent reaction intermediates to assess the structural and electronic basis of enzyme function on a detailed molecular level.