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Structure and Function of AMPA subtype ionotropic glutamate receptors

AMPA 亚型离子型谷氨酸受体的结构和功能

基本信息

批准号：
10197227
负责人：
MARIA G KURNIKOVA
金额：
$ 50.13万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-30 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10197227
关键词：
AMPA Receptors Adamantane Alzheimer&apos s Disease Animal Model Antiepileptic Agents Behavior Binding Sites Biochemical Biophysics Cell Line Cells Color Complement Complex Cryoelectron Microscopy Crystallization Dementia Development Disease Drug Design Drug Targeting Electrophysiology (science)Epilepsy Epileptogenesis Eukaryotic Cell Family Fluorescence Future Glutamate Receptor Goals Homo Individual Insecta Ion Channel Ischemia Knowledge Learning Length Maps Mediating Memory Mental disorders Methodology Methods Modeling Modernization Molecular Molecular Structure Monitor Mutagenesis Neuraxis Neurodegenerative Disorders Pharmaceutical Preparations Pharmacotherapy Play Polyamines Positioning Attribute Primary Lateral Sclerosis Proteins Protocols documentation Regulation Research Resolution Seizures Structural Models Structure Techniques Testing Therapeutic Toxin Transfection Viral base biophysical model cognitive process design drug candidate drug development drug testing experimental study expression vector human disease human model improved in silico inhibitor/antagonist ion channel blocker kinetic model molecular dynamics molecular modeling nervous system disorder neurotransmission next generation novel therapeutics patch clamp promoter receptor expression receptor structure function scaffold small molecule

项目摘要

AMPA receptors mediate fast excitatory neurotransmission, contribute to high cognitive processes such as learning and memory and are implicated in numerous psychiatric and neurodegenerative diseases. In particular, AMPA receptors play a key role in epileptogenesis and seizure spread and, thus, have recently emerged as one of the most promising targets for epilepsy therapy. However, development of drugs targeting AMPA receptors has been stalled because of the lack of knowledge about AMPA receptor structure and function. For example, only structures of homotetrameric intact AMPA receptors have been determined, while the overwhelming majority of AMPA receptors in the central nervous system are heterotetramers. A number of noncompetitive inhibitors and ion channel blockers have been identified as promising candidates for drug development but structural mechanisms of their action on AMPA receptors remain largely unexplored. This missing information is absolutely critical for the future structure-based rational drug design. We plan to study structure and function of AMPA receptors using a combination of biophysical and biochemical approaches, including modern crystallographic and cryo-electron microscopy (cryo-EM) techniques, fluorescence-based methods, electrophysiology, kinetic and molecular modeling. Our specific aims are to (1) obtain structures of heteromeric AMPA receptors, (2) establish the molecular mechanism of noncompetitive inhibition, and (3) build a structural model of ion channel block. To reach our goals, we will optimize AMPA receptor constructs for crystallization and cryo-EM experiments, develop protocols of their expression and purification and solve structures of heterotetrameric AMPA receptors and AMPA receptors in complex with noncompetitive inhibitors and ion channel blockers. To improve our structural models, we will use new methods of structural refinement combined with molecular dynamics (MD) simulations. We will also test our models using a combination of experimental and in silico mutagenesis, whole-cell patch-clamp recordings and MD simulations. To understand the molecular mechanisms of AMPA receptor heteromeric assembly, noncompetitive inhibition and ion channel block, we will perform extensive MD simulations of homo- and heteromeric AMPA receptors in different activation states and in the presence or absence of noncompetitive inhibitors and ion channel blockers. We will combine the results of structural, computational, functional and mutagenesis experiments to propose molecular models of AMPA receptor heteromeric assembly, noncompetitive inhibition and ion channel block. Reaching our research goals will provide molecular level knowledge essential to greatly facilitate design of new molecules that will have a potential to become safe and more efficacious drugs to treat epilepsy and other disorders related to excitatory neurotransmission.

AMPA受体介导快速兴奋性神经传递，有助于高认知过程，如