HIGH RESOLUTION CRYSTALLOGRAPHY OF HEME ENZYMES WITH SUBSTRATE-LINKED SENSITIZER

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。目前，人们对底物进入途径和底物与P450结合相关的构象变化非常感兴趣。为了充分分析这些特定的相互作用，已经开发了底物连接敏化剂(SLS)探针。到目前为止，已经解决了Ru三联吡啶探针与P450 cam结合的1.55和1.65结构，以及荧光基探针与P450 cam结合的2.2、1.8和1.45结构。随着设计过程继续优化用于光谱研究和抑制剂识别的SLS探测器，将在第二代探测器中探索替代荧光团。因此，这项提案中的高分辨率结晶学研究将大大有助于开发酶特异性探针的长期目标，用于分析分子识别的复杂机制，以及快速筛选具有生物医学意义的重要酶的抑制剂，如细胞色素P450。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Currently there is substantial interest in the substrate access pathway and the conformational changes associated with substrate binding to P450s. To fully analyze these specific interactions, substrate-linked sensitizer (SLS) probes have been developed. To date, 1.55 and 1.65 ¿¿structures of ruthenium tris-bipyridyl probes bound to P450cam have been solved, along with 2.2, 1.8, and 1.45 ¿¿structures of fluorescent-based probes bound to P450cam. As the design process continues to optimize the SLS probes for spectroscopic studies and inhibitor identification, alternative fluorophores will be explored in a second generation of probes. Thus, the high-resolution crystallographic studies in this proposal will significantly aid in the long-term goal of developing enzyme-specific probes for analyzing the complex mechanisms of molecular recognition as well as for rapid screening of inhibitors of bio-medically important enzymes such as cytochrome P450s.