Probing the Mechanism and Structure-Function Relations in Phenylalanine Hydroxylase

苯丙氨酸羟化酶的作用机制及构效关系探讨

• 批准号: 0434637
• 负责人: Mahdi Abu-Omar
• 金额: $ 24.98万
• 依托单位: Purdue University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0434637&HistoricalAwards=false
• 关键词: Probing; Mechanism; Structure; Function; Relations

This award by the Inorganic, Bioinorganic and Organometallic Chemistry program supports research by Professor Mahdi Abu-Omar of the University of California at Los Angeles to study the enzyme, phenylalanine hydroxylase from chromobacterium violaceum (cPAH). A genetic deficiency in the homologous human enzyme, hPAH, is responsible for the disease phenylketonuria. This research involves: 1) determining the rate-limiting step in the catalytic reaction of PAH by measuring 18O kinetic isotope effects; 2) trapping and characterizing intermediates involved in oxygen activation in PAH by kinetic and spectroscopic methods; 3) understanding the active site architecture and the origin of the substrate selectivity through site-specific mutants of the enzyme; and 4) establishing structure-function relationships by creating and exploring the reactivity of new PAH chimera composed of the regulatory domain of hPAH and the catalytic domain of cPAH.The function of an important enzyme, phenylalanine hydroxylase, which synthesizes an essential amino acid, tyrosine that is further metabolized to important neurotransmitters will be studied. The work will focus on how oxygen is activated at its iron-containing active site.

该奖项获得了无机，生物有机和有机金属化学计划的支持，支持了加利福尼亚大学洛杉矶分校的Mahdi Abu-Omar教授的研究，以研究紫collomobacterium collomobacterium utaceum（CPAH）的苯丙氨酸羟化酶。 同源人酶HPAH的遗传缺陷是疾病苯酮尿症的原因。 这项研究涉及：1）通过测量18o动力学同位素效应来确定PAH催化反应的速率限制步骤； 2）通过动力学和光谱方法捕获和表征PAH中参与氧气激活的中间体； 3）通过酶的位点特异性突变体了解活性位点结构和底物选择性的起源； 4）通过建立和探索由HPAH调节域和CPAH的催化域组成的新PAH嵌合体的反应性来建立结构功能关系。 这项工作将集中于如何在其含铁的活性位点激活氧气。