ULTRA HIGH RESOLUTION STRUCTURE OF A NOVEL TRANSFERASE ENZYME

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。这种新型转移酶可催化小基团转移反应。我们利用MAD相位技术确定了一种新型转移酶的晶体结构，并将其细化到1.5¿¿分辨率。对结构的分析揭示了这类酶迄今为止未知的八聚体结构，在单个亚基之间具有有趣的相互作用。值得注意的是，该酶表现出复杂的反应动力学。因此，酶的超高分辨率对于正确理解反应机理是有价值的。在这项研究中，我们重点收集了具有共价结合抑制剂的酶的数据。该配合物结晶为三斜单晶胞。晶体衍射优于1.0¿¿，我们收集了1.0¿¿的数据集，总体完成率为80%，Rsym为3.8%。最高分辨率壳体（1.04 ~ 1.00 ~ 1.00）的I/sig(I)为4.3 sigma。

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. This novel transferase enzyme catalyzes the transfer reactions of small groups. We have determined the crystal structure of a novel transferase enzyme using MAD phasing techniques, and refined it to 1.5 ¿¿resolution. Analysis of the structure revealed a hitherto unknown octameric structure of this class of enzymes with interesting interactions between the individual subunits. It is also noteworthy that this enzyme shows complex reaction kinetics. Therefore, an ultra high resolution of the enzyme is valuable for a proper understanding of the reaction mechanism. In this study we focused on the data collection of the enzyme with a covalently bound inhibitor. The complex crystallized with a triclinic unit cell. The crystals diffracted to better than 1.0 ¿¿and we collected a 1.0 ¿¿data set with 80% overall completion and an Rsym of 3.8%. The I/sig(I) for the highest resolution shell (1.04 ¿¿ 1.00 ¿¿ is 4.3 sigma.