Structural Studies of Vesicular Monoamine Transporters

囊泡单胺转运蛋白的结构研究

基本信息

批准号：
8528735
负责人：
SATINDER Kaur SINGH
金额：
$ 19.98万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
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/8528735
关键词：
1-Methyl-4-phenylpyridinium Alzheimer&apos 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

项目摘要

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.

描述（由申请人提供）：囊泡单胺转运蛋白 2 (VMAT2) 是一种整合膜蛋白，通过补充突触前囊泡中耗尽的神经递质储备，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个管腔质子的流出耦合起来。底物包括血清素、多巴胺、去甲肾上腺素、肾上腺素和组胺。 VMAT2 的重要性通过其在突触小泡内浓缩其底物的惊人水平 (~500 mM) 及其在隔离神经元细胞质中的几种有毒化合物方面的积极作用而得到强调。目前的处方疗法包括抑制剂利血平[RES]和丁苯那嗪[TBZ]。 RES 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舞蹈症中的失控运动和抽动秽语综合征中的抽动。尽管具有如此的临床和药理学意义，但人们对转运、离子依赖性、底物识别和抑制的结构基础知之甚少。 VMAT2 晶体结构将提供关键信息，并可能彻底改变该领域。该提案的目标是收集实现这一目标所需的初步数据：表达、纯化和获得真核 VMAT2 同源物的衍射质量晶体。