P2 - Molecular Mechanisms in Monoamine Transporter Function

P2 - 单胺转运蛋白功能的分子机制

• 批准号: 8376685
• 负责人: Ulrik Gether
• 金额: $ 12.5万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8376685
• 关键词: Acute; Alanine; Amphetamines; Behavioral; Binding; Brain; C-terminal; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium/calmodulin-dependent protein kinase; Carrier Proteins; Cell membrane; Cell physiology; Cells; Cocaine; Cocaine Abuse; Corpus striatum structure; Data; Development; Distal; Dopamine; Dose; Ensure; Environment; Exposure to; Extracellular Space; Family; Functional disorder; Future; Goals; HIV-1; Investigation; Ions; Knock-in Mouse; Knockout Mice; Laboratories; Libraries; Lipids; Mediating; Membrane; Membrane Microdomains; Mental disorders; Molecular; Movement; Mus; Mutate; Neurons; Neurotransmitters; PDZ protein; Participant; Peptides; Pharmaceutical Preparations; Phenotype; Phosphotransferases; Physiological; Principal Investigator; Process; Property; Protein Kinase; Proteins; Role; Signal Transduction; Societies; Sodium; Structure; Tertiary Protein Structure; Work; addiction; base; dopamine transporter; drug abuse therapy; in vivo; insight; intravenous administration; monoamine; multidisciplinary; mutant; novel therapeutics; presynaptic; protein protein interaction; psychostimulant; response; socioeconomics; stimulant abuse; symporter; tat Protein; trafficking

The dopamine transporter (DAT) is the primary target for the abused psychostimulants such as amphetamines (AMPHs) and cocaine. DAT belongs to the family of NeurotransmitterSodium: Symporters (NSS) and Is responsible for rapid clearance of dopamine from the extracellular space. It is our long term goal to understand the molecular and cellular processes governing the activity and availability of DAT in the presynaptic membrane and how these processes are altered in diseased states involving dysfunction of dopaminergic signaling. To perform its function, DAT participates in a network of protein-protein interactions and protein-llpid interactions that ensure proper trafficking, localization and catalytic activity of the transporter. We hypothesize that interaction ofthe DAT C-terminus with Ca2+/calmodulin dependent protein kinase Ila (CaMKIlo) and with PDZ (PSD-95/Discs-large/ZO-1 homology) domain proteins Is of central mportance for these processes. In knock-in mice expressing a DAT mutant incapable of forming PDZ interactions we observe a dramatic decrease in axonal and dendritic localization ofthe transporter. Our data suggest that this phenotype is not caused by disrupting the interaction with PICKI (protein interacting with C kinase-1), the so far only PDZ protein known to Interact with DAT, but rather the interaction with yet unidentified PDZ proteins. Instead we hypothesize that PICKI binding regulates DAT electrogenic properties. An in vivo role of C-terminal protein-protein interactions in mediating AMPH action is supported by the ability of a C-terminal membrane permeable DAT peptide to reduce the locomotor response to acute AMPH in mice. We propose therefore i) to determine the role of PDZ interactions in governing axonal and dendritic localization of DAT, and to examine the physiological consequences of disrupting these interactions; 11) to investigate the role of PICKI in regulating DAT ion conductances and AMPH-induced dopamine efflux; and iii) to determine the role, in vivo, that binding of CaMKIIa and PICKI to the DAT C- terminus has in establishing the behavioral effects of AMPH. These investigations should contribute significantly to future development of better therapies for drug abuse and perhaps psychiatric disorders. RELEVANCE (See instmctions): The abuse of cocaine and amphetamine causes profound socio-economic problems in the society. However, we know still little about the mechanisms in the brain that are responsible for the development of addiction to these drugs. Insight into the molecular mechanisms governing the activity and availability of their primary target, the dopamine transporter, should prove Instrumental in our search for new therapeutic strategies.

多巴胺转运蛋白（DAT）是滥用心理刺激物（例如）的主要目标 苯丙胺（AMPHS）和可卡因。 DAT属于神经递质的家族：隔离器 （NSS），负责从细胞外空间快速清除多巴胺。这是我们的长期 了解有关DAT活动和可用性的分子和细胞过程 突触前膜以及如何在涉及功能障碍的病态中改变这些过程 多巴胺能信号传导。为了执行其功能，DAT参与了蛋白质 - 蛋白质相互作用的网络 和蛋白质差异相互作用，以确保适当的运输，定位和催化活性 转运蛋白。我们假设Dat C末端与Ca2+/钙调蛋白依赖性蛋白的相互作用 激酶ILA（CAMKILO）和PDZ（PSD-95/DISCS-LARGE/ZO-1同源性）域蛋白质是中心的 这些过程的忠诚度。在表达无法形成PDZ的DAT突变体的敲入小鼠中 相互作用我们观察到转运蛋白的轴突和树突定位急剧下降。我们的数据 表明这种表型不是由于破坏与Picki的相互作用而引起的（蛋白质与C相互作用 激酶-1），迄今为止仅PDZ蛋白已知与DAT相互作用，而是与 身份不明的PDZ蛋白。相反，我们假设Picki结合调节DAT电源 特性。支持C末端蛋白质蛋白相互作用在介导AMPH作用中的体内作用 通过C末端膜可渗透DAT肽的能力减少对急性的运动反应 小鼠的Amph。因此，我们建议i）确定pdz相互作用在轴突和 DAT的树枝状定位，并检查破坏这些的生理后果 互动； 11）调查Picki在调节DAT离子电导和AMPH诱导的作用 多巴胺外排； iii）为了确定camkiia和picki与dat c-的结合的作用 末端在建立AMPH的行为效应方面具有。这些调查应该做出贡献 重要的是，未来对药物滥用和精神疾病的更好疗法的发展。 相关性（请参阅Instmctions）： 可卡因和苯丙胺的滥用会导致社会上的深刻社会经济问题。然而， 我们对大脑中造成成瘾发展的大脑机制知之甚少 这些药物。深入了解其主要的活性和可用性的分子机制 多巴胺转运蛋白的靶标应证明在我们寻求新的治疗策略方面具有重要作用。