OLIGOMERIZATION STATE OF THE MONOAMINE OXIDASES

单胺氧化酶的低聚状态

• 批准号: 8363971
• 负责人: PETER P BORBAT
• 金额: $ 0.05万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363971
• 关键词: Active Sites; Amines; Binding; Data; Enzymes; Funding; Grant; Human; Measurement; Membrane; Monoamine Oxidase; Monoamine Oxidase A; Mutation; National Center for Research Resources; Outer Mitochondrial Membrane; Pargyline; Play; Positioning Attribute; Principal Investigator; Rattus; Research; Research Infrastructure; Resources; Role; Sampling; Source; Spin Labels; Structure; Study models; Technology; Testing; United States National Institutes of Health; analog; cost; inhibitor/antagonist; mutant; neurotransmitter metabolism; quantum

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. Monoamine oxidases ( A & B) are outer mitochondrial membrane-bound enzymes that play important roles in amine neurotransmitter metabolism. Sequence and modeling studies suggest human MAO A differs from all other mammalian MAO A in that it is monomeric rather than dimeric in the outer membrane due to a Glu151Lys mutation. Crystal structures of purified human and rat MAO A provide additional support for this hypothesis. To provide a direct experimental test, analogues of the inhibitor pargyline with a nitroxide spin label on the para position will be synthesized and covalently incorporated into the active sites of human and rat MAO A, the Lys151Glu mutant of human MAO A, and human MAO B. Structural data of the dimeric form of crystalline human MAO B show the distance between the active site cavities is 42 ¿. This distance is readily determined by Double Quantum Coherence EPR measurements on these samples. The dipolar spectral data will provide distance information that will determine if these enzyme species are in their dimeric or monomeric forms in the mitochondrial outer membrane.

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 该子项目的主要支持。 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源的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。 单胺氧化酶 (A 和 B) 是线粒体外膜结合酶，在胺神经递质代谢中发挥重要作用。序列和建模研究表明，人类 MAO A 与所有其他哺乳动物 MAO A 不同，因为它在外膜中是单体而非二聚体。由于 Glu151Lys 突变，纯化的人和大鼠 MAO A 的晶体结构为这一假设提供了额外的支持。对位上的硝基氧自旋标签将被合成并共价掺入人和大鼠 MAO A、人 MAO A 的 Lys151Glu 突变体和人 MAO B 的活性位点。结晶人 MAO B 二聚体形式的结构数据显示活性位点腔之间的距离为 42 ¿该距离很容易通过对这些样品进行双量子相干 EPR 测量来确定。偶极光谱数据将提供距离信息，从而确定这些酶种类在线粒体外膜中是否处于二聚体或单体形式。