X-RAY CRYSTALLOGRAPHIC STUDIES OF METAL-REQUIRING ENZYMES

这个子项目是利用资源的许多研究子项目之一 由NIH/NCRR资助的中心拨款提供。次级项目的主要支助 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们在NE-CAT的工作主要集中在一些金属酶系统，这些系统的晶体质量和尺寸使他们的研究在其他同步辐射源上变得困难。该实验室的一个关键项目集中在双核锰金属酶脱氢酶，以及共享脱氢酶折叠的酶，如锌酶人类组蛋白脱乙酰基酶-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，其催化革兰氏阴性菌中脂质A生物合成的第一个关键步骤。这些金属酶的比较，最终将揭示的化学和结构的基础上的分歧进化的金属离子的特异性和化学计量的酶折叠。此外，我们在NE-CAT的工作集中在萜类环化酶上。这些酶需要金属，催化生物学中最复杂的有机化学反应。 具体而言，我们的工作集中在倍半萜环化酶，催化环化反应，涉及底物法呢基二磷酸或香叶基香叶基二磷酸：epi-isozizaene合酶和柯巴基二磷酸合酶。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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 substrates farnesyl diphosphate or geranylgeranyl diphosphate: epi-isozizaene synthase and copalyl diphosphate synthase.