Forefront X-Ray Crystallography of Enzymes by using the third generation synchrotron radiation

1.时间分辨x射线结晶学--两种动力学结晶学技术的结合--衬底连续流动和劳埃衍射测量，使我们能够在反应进行之前捕捉到跃迁结构。在反应开始前，我们测定了酶-底物复合体的晶体结构：托品酮还原酶-II-NADPH-托品诺酮。很复杂。本研究中的酶-底物复合体与我们先前研究中的酶-产物复合体的结构比较表明，底物托品酮是相对于产物旋转的，从而使活性中心的空间组织有利于反应的进行。2.超高分辨X射线晶体学在原子分辨率下解析了天然状态和两种半乳糖醛酸络合态的紫草内切多聚半乳糖醛酸酶的晶体结构。3.困难分析的X射线晶体结构测定我们测定了玉米丙酮酸磷酸二激酶的晶体结构，这是一种含有钙调蛋白结合域的Marcks多肽，它与钙(2)-钙调素形成络合物，与钙(2)-钙调素形成络合物，与钙(2)钙调素形成络合物。

1.time-resolved x-ray crystallography A combination of two kinetic crystallographic techniques, a continuous flow of the substrates and Laue diffraction measurement, enabled us to capture the transit structures prior to the reaction proceeding. We determined the crystal stuructre of enzyme-substrate complex prior to reaction initiation: tropinone reductase-II-NADPH-tropinone. complex. A structural comparison of the enzyme-substrate complex elucidated in this study with the enzyme-product complex in our previous study indicated that one of the substrates, tropinone, is rotated relative to the product so as to make the spatial organization in the active site favorable for the reaction to proceed.2.ultra-high-resolution X-ray crystallography Crystal structures of endopolygalacturonase from Stereum purpureum were solved in native and two galacturonic acid complex states at atomic resolution. The active-site architecture of the complexes provides insight into the mechanism of inverting glycosyl hydrolases and also sheds light on the basis of the differences between the family 28 and the other inverting glycosyl hydrolases.3.X-ray crystal structure determination with difficult analysis We determined crystal structure of maize pyruvate phosphate dikinase, a MARCKS peptide containing the calmodulin-binding domain in complex with Ca(2+)-calmodulin, a myristoylated CAP-23/NAP-22N-terminal domain complexed with Ca(2+)calmodulin, and the PsbP protein of photosystem II from Nicotiana tabacum.