ULTRA HIGH RESOLUTION STRUCTURE OF A NOVEL TRANSFERASE ENZYME

新型转移酶的超高分辨率结构

• 批准号: 7370678
• 负责人: IRIMPAN I MATHEWS
• 金额: $ 0.09万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7370678
• 关键词: ULTRA; RESOLUTION; STRUCTURE; NOVEL; TRANSFERASE

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.

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。这种新型的转移酶催化小分子基团的转移反应。我们用MAD相变技术测定了一种新型转移酶的晶体结构，并将其提纯到1.5°分辨率。对结构的分析揭示了这类酶的一种迄今未知的八聚体结构，各个亚基之间存在有趣的相互作用。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。该络合物结晶为三斜晶胞。晶体的衍射率高于1.0°，我们收集了一个总完成率为80%，Rsym为3.8%的1.0°数据集。最高分辨率壳层(1.04)的i/sig(I)为4.3西格玛。