X-RAY CRYSTALLOGRAPHIC STUDIES OF METAL-REQUIRING ENZYMES

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们在 NE-CAT 的工作重点是许多金属酶系统，这些系统的晶体质量和尺寸使得在其他同步加速器源上进行研究变得困难。该实验室的一个关键项目侧重于双核锰金属酶精氨酸酶，以及共享精氨酸酶折叠的酶，例如锌酶、人组蛋白脱乙酰酶 8 和细菌乙酰多胺酰胺水解酶。这项工作还包括来自小肠结肠炎耶尔森氏菌的锌依赖性脱乙酰酶 LpxC，它催化革兰氏阴性细菌中脂质 A 生物合成的第一个关键步骤。这些金属酶的比较将最终揭示精氨酸酶折叠中金属离子特异性和化学计量的不同进化的化学和结构基础。此外，我们在 NE-CAT 的工作重点是萜类环化酶。这些是需要金属的酶，可以催化生物学中最复杂的有机化学反应。 具体来说，我们的工作重点是倍半萜环化酶，其催化涉及底物法尼基二磷酸的环化反应：δ杜松烯合酶和表异齐烯合酶的位点特异性突变体。

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. Our work at NE-CAT has focused on a number of metalloenzyme systems for which crystal quality and size make their study difficult at other synchrotron sources. A key project in the lab focuses on the binuclear manganese metalloezyme arginase, as well as enzymes that share the arginase fold such as the zinc enzymes human histone deacetylase-8 and bacterial acetylpolyamine amidohydrolase. This work additionally includes the zinc-dependent deacetylase LpxC from Yersinia enterocolitica, which catalyzes the first committed step of lipid A biosynthesis in Gram-negative bacteria. Comparisons of these metalloenzymes will ultimately reveal the chemical and structural basis for the divergent evolution of metal ion specificity and stoichiometry in the arginase fold. Additionally, our work at NE-CAT has focused on terpenoid cyclases. These are metal-requiring enzymes that catalyze the most complex organic chemical reactions in biology. Specifically, our work has focused on sesquiterpene cyclases that catalyze cyclization reactions involving the substrate farnesyl diphosphate: delta cadinene synthase and site-specific mutants of epi-isozizaene synthase.