Structural Studies of Vesicular Monoamine Transporters

囊泡单胺转运蛋白的结构研究

• 批准号: 8360957
• 负责人: SATINDER Kaur SINGH
• 金额: $ 23.25万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-10 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360957
• 关键词: 1-Methyl-4-phenylpyridinium; Alzheimer' s Disease; Amines; Amphetamines; Antibodies; Area; Binding; Bipolar Disorder; Birth; Brain; Brain imaging; Cell membrane; Cells; Cholesterol; Clinical; Cocaine; Collaborations; Coupled; Crystallization; Cytoplasm; Data; Dependence; Detergents; Development; Disease; Dopamine; Drug Addiction; Drug Design; Enhancers; Epinephrine; Fluorescence; Funding; Genetic Polymorphism; Gilles de la Tourette syndrome; Glycerol; Goals; Grant; Heterogeneity; Histamine; Homologous Gene; Huntington Disease; Hypertension; Insecta; Integral Membrane Protein; Ion Transport; Knockout Mice; Laboratories; Length; Link; Lipids; Mammals; Membrane; Membrane Proteins; Molecular; Molecular Conformation; Molecular Sieve Chromatography; Monitor; Mood Disorders; Movement; Muramidase; Mus; Mutagenesis; Neuraxis; Neurons; Neurotoxins; Neurotransmitters; Norepinephrine; Parkinson Disease; Parkinsonian Disorders; Patients; Phase; Play; Poison; Process; Protein Isoforms; Proteins; Protons; Psychotic Disorders; Publishing; Rattus; Reducing Agents; Research; Reserpine; Resources; Role; Schizophrenia; Serotonin; Shapes; Signal Transduction; Sodium; Spodoptera frugiperda; Structure; Substantia nigra structure; Surface; Synapses; Synaptic Cleft; Synaptic Transmission; Synaptic Vesicles; Tardive Dyskinesia; Temperature; Testing; Tetrabenazine; Therapeutic; Vesicle; Work; Yeasts; analog; antiport; base; density; design; dihydrotetrabenazine; directed evolution; high risk; inhibitor/antagonist; insight; interest; monoamine; mutant; neuropsychiatry; neurotransmitter release; overexpression; presynaptic; radioligand; response; stability testing; success; symporter; transport inhibitor; uptake; vacuolar H+-ATPase; vesicular monoamine transporter; vesicular monoamine transporter 1; vesicular monoamine transporter 2

DESCRIPTION (provided by applicant): Vesicular monoamine transporter 2 (VMAT2) is an integral membrane protein that plays a critical role in aminergic synaptic transmission by replenishing depleted neurotransmitter stores in presynaptic vesicles. It works by harnessing a pre-existing proton gradient generated by V-type ATPase to couple the uptake of one cytoplasmic cationic amine with the efflux of two luminal protons. Substrates include serotonin, dopamine, norepinephrine, epinephrine, and histamine. The significance of VMAT2 is underscored by the phenomenal levels (~500 mM) to which it concentrates its substrates inside the synaptic vesicle and by its active role in sequestering several toxic compounds from the neuronal cytoplasm. Currently-prescribed therapeutics include the inhibitors reserpine [RES] and tetrabenazine [TBZ]. RES is used as a second-line agent to treat hypertension and psychosis; TBZ is approved to treat the uncontrolled movements in Huntington's disease and the tics in Tourette's syndrome. Despite such clinical and pharmacological significance, very little is known about the structural basis for transport, ion-dependence, substrate recognition, and inhibition. A VMAT2 crystal structure would provide the crucial information and likely revolutionize the field. The goal of this proposal is to gather the preliminary data required for such a pursuit: to express, purify, and obtain diffraction-quality crystals of a eukaryotic VMAT2 homologue. PUBLIC HEALTH RELEVANCE: Vesicular monoamine transporter 2 (VMAT2) plays a critical role in the overall process of synaptic transmission by replenishing depleted monoamine stores in synaptic vesicles. It is the target of the noncompetitive inhibitor tetrabenazine, clinically usd for symptomatic treatment of hyperkinetic disorders such as Huntington's disease, Tourette's syndrome, and tardive dyskinesia. Thus, understanding how this integral membrane works at the atomic level is a crucial goal.

描述（由申请人提供）：囊泡单胺转运蛋白2（VMAT 2）是一种整合的膜蛋白，通过补充突触前囊泡中耗尽的神经递质储存，在胺能突触传递中发挥关键作用。它的工作原理是利用V型ATP酶产生的预先存在的质子梯度，将一种细胞质阳离子胺的摄取与两种腔质子的流出偶联。底物包括血清素、多巴胺、去甲肾上腺素、肾上腺素和组胺。VMAT 2的重要性通过其将其底物集中在突触囊泡内的显著水平（~500 mM）以及其在从神经元细胞质中隔离几种有毒化合物中的积极作用来强调。目前处方的治疗剂包括抑制剂利血平[RES]和丁苯那嗪[TBZ]。RES被用作治疗高血压和精神病的二线药物; TBZ被批准用于治疗亨廷顿病中的不受控制的运动和抽动秽语综合征中的抽搐。尽管有这样的临床和药理学意义，但对转运、离子依赖性、底物识别和抑制的结构基础知之甚少。VMAT 2晶体结构将提供关键信息，并可能使该领域发生革命性变化。该提案的目标是收集这种追求所需的初步数据：表达，纯化和获得真核VMAT 2同源物的衍射质量晶体。 公共卫生关系：囊泡单胺转运体2（VMAT 2）通过补充突触囊泡中耗尽的单胺储存在突触传递的整个过程中起着关键作用。它是非竞争性抑制剂丁苯那嗪的靶点，临床上用于对症治疗运动过度障碍，如亨廷顿病、图雷特综合征和迟发性运动障碍。因此，了解这种完整的膜如何在原子水平上工作是一个至关重要的目标。