STRUCTURE-ACTIVITY RELATIONSHIP STUDIES OF DOPAMINE BETA-MONOOXYGENASE

多巴胺β-单加氧酶的构效关系研究

• 批准号: 8359663
• 负责人: KANDATEGE WIMALASENA
• 金额: $ 9.9万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8359663
• 关键词: Active Sites; Biochemical; Catecholamines; Cocaine Dependence; Congestive Heart Failure; Copper; Development; Disease; Dopamine-beta-monooxygenase; Enzymes; Etiology; Funding; Grant; Human; Hypertension; Maintenance; Mixed Function Oxygenases; Molecular; National Center for Research Resources; Neurotransmitters; Play; Principal Investigator; Proteins; Recombinants; Regulation; Research; Research Infrastructure; Resources; Role; Site-Directed Mutagenesis; Source; Structure-Activity Relationship; Sympathetic Nervous System; System; Therapeutic; United States National Institutes of Health; chemical kinetics; cost; in vivo; mutant; neurotransmitter biosynthesis; protein structure function

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. The copper enzyme, dopamine beta-monooxygenase (DbM; E.C. 1.14.17.1), plays a central role in catecholamine neurotransmitter biosynthesis. Consequently, the chemical and the kinetic mechanisms of DbM have been extensively studied during the last five decades. However, the correlation of the experimental findings with the structural parameters of D-beta-M is lagging behind due to the lack of structural and molecular details of the active site of the enzyme. The overall objective of the proposed studies is to express human D-beta-M in a system suitable for site-directed mutagenesis studies and to carry out systematic biochemical, biophysical, and structural and mechanistic studies using purified recombinant wild type and mutant proteins. D-beta-M has long been recognized as an important target for the modulation of sympathetic nervous system activity for therapeutic purposes due to its central role in the biosynthesis of the neurotransmitter NE. For example, the vital role of the sympathetic nervous system and its neurotransmitter NE in the development and maintenance of hypertension and congestive heart failure have been extensively studied. Recent studies have also shown that the regulation of DbM activity in vivo may have beneficial effects on the treatment of cocaine addiction. Therefore, better understanding of the structure-activity relationship of DbM at the molecular level will be important in determining the etiology of these diseases and eventual development of effective therapeutics. In addition, DbM is prototypical of a large group of relatively more specific non-heme monooxygenases and the details of its catalytic mechanism.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 铜酶，多巴胺β-单加氧酶（DbM；E.C. 1.14.17.1），在儿茶酚胺神经递质生物合成中发挥核心作用。因此，在过去的五年中，DbM 的化学和动力学机制得到了广泛的研究。然而，由于缺乏酶活性位点的结构和分子细节，实验结果与 D-β-M 结构参数的相关性滞后。拟议研究的总体目标是在适合定点诱变研究的系统中表达人类 D-β-M，并使用纯化的重组野生型和突变蛋白进行系统的生化、生物物理以及结构和机制研究。由于 D-β-M 在神经递质 NE 生物合成中的核心作用，D-β-M 长期以来被认为是调节交感神经系统活性以达到治疗目的的重要靶标。例如，交感神经系统及其神经递质NE在高血压和充血性心力衰竭的发生和维持中的重要作用已被广泛研究。最近的研究还表明，体内 DbM 活性的调节可能对可卡因成瘾的治疗产生有益的影响。因此，在分子水平上更好地了解 DbM 的结构-活性关系对于确定这些疾病的病因和最终开发有效的治疗方法非常重要。此外，DbM 是一大类相对更具特异性的非血红素单加氧酶的原型及其催化机制的细节。