CRYSTAL STRUCTURES OF ACYL-COA CARBOXYLASE AS TARGETS OF CANCER AND OBESITY THER

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 乙酰辅酶A羧化酶(ACCase)，如乙酰辅酶A羧基酶(ACC)或丙基辅酶A羧基酶(PCC)，催化乙酰辅酶A和丙基辅酶A的羧化反应，生成丙二酰辅酶A和甲基丙二酰辅酶A。这种羧化反应是最重要的代谢调节检查点之一，它将乙酰辅酶A和丙基辅酶A委托给脂肪酸的生物合成。因此，ACC和PCC是肥胖症、癌症和糖尿病治疗的极有希望的靶点。核心催化亚基PCCB和ACCB是360 kDa的同源六聚体，催化生物素-CO2和酰辅酶A之间的转羧化。我们解决了PCCB六聚体的脱脂晶体结构到2.0°。为了了解ACCase的分子识别，生长了野生型和突变型ACCB和PCCB的衍射级晶体，并在室内将其指标化到3.5？4°，包括载脂蛋白、二元(酶-生物素)和三级(酶-生物素-酰基辅酶A)晶体。一些晶体形式需要重原子方法(MIR和MAD)来解决问题。SSRL的光束时间将是我们成功解决ACCase高分辨率结构的关键。结合突变数据，这些结构将揭示底物识别的分子基础，从而确定未来癌症和肥胖症治疗的药物设计目标。

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. Acyl-coenzyme A carboxylases (ACCase), such as acetyl-CoA carboxylase (ACC) or propanyl-CoA carboxylase (PCC), catalyze the carboxylation of acetyl- and propanyl-CoA to provide malonyl- and methylmalonyl-CoA. This carboxylation reaction is one of the most important metabolic regulation checkpoints by committing acetyl-CoA and propanyl?CoA to the biosynthesis of fatty acids. ACC and PCC are therefore a highly promising target for obesity, cancer and diabetes therapy. The core catalytic subunits, pccB and accB, are 360 kDa homo-hexamers, catalyzing the transcarboxylation between biotin-CO2 and acyl-CoA. We solved the apo crystal structures of pccB hexamer to 2.0 ¿. To understand the molecular recognition of ACCase, difffration-quality crystals of both wild type and mutant accB and pccB were grown, diffracted and indexed in-house to 3.5?4 ¿, including apo, binary (enzyme-biotin) and tertiary (enzyme-biotin-acyl CoA) crystals. Some crystal forms will require heavy atom methods (MIR and MAD) for solutions. Beamtime at SSRL will be vital for our success to solve high resolution structures of ACCase. The structures, combined with mutagenesis data, will shed light on the molecular basis of substrate recognition, resulting in the identification of future drug design targets for cancer and obesity therapy.