Engineering a Transmitter Binding Site

设计发射器绑定位点

• 批准号: 8074901
• 负责人: Anthony L Auerbach
• 金额: $ 33.63万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8074901
• 关键词: Address; Binding; Binding Sites; Biological Models; Biology; Cholinergic Receptors; Complex; Coupling; Development; Drug effect disorder; Electrophysiology (science); Engineering; Equilibrium; Event; Family; Health; Ions; Kinetics; Knowledge; Ligand Binding; Ligands; Maps; Measures; Medicine; Molecular; Molecular Conformation; Mono-S; Mutation; Nature; Neurotransmitters; Pharmaceutical Preparations; Pharmacologic Substance; Physiological; Proteins; Shapes; Side; Structure; base; design; drug discovery; improved; mutant; neuromuscular; protein complex; protein structure function; receptor; research study; response; small molecule

DESCRIPTION (provided by applicant): A fundamental principle of biomedicine is that small molecules can bind to specific receptors to trigger physiological responses. We seek to understand the energetic nature of the molecular events that occur at the neurotransmitter binding sites of the neuromuscular acetylcholine receptor when this channel 'gates' between non-conducting and ion-conducting conformations. This knowledge will help us understand the biophysical mechanisms of ligand-protein complexes, and will advance our ability to design new drugs for nicotinic (and other) receptors. Structure-based drug discovery should incorporate the fact that drug action depends on the differential binding of ligands to inactive vs. active conformations of a receptor. To address this point we will use single-channel electrophysiology and kinetic analysis to study unliganded gating, which will allow us to measure all of the salient activation equilibrium constants and to ascertain the energetic contributions of the side chains at each of the two transmitter binding sites. We will also estimate differential binding energies for small ligand probes, in both wild type and mutant receptors. Eventually, this knowledge will be used to engineer acetylcholine receptors that respond predictably to arbitrary ligands. PUBLIC HEALTH RELEVANCE: The experiments outlined in this project establish the fundamental principles for engineering a protein to respond in predictable ways to specific drugs. These principles can be applied to the development of new pharmaceuticals, and to understanding the mechanism by which a drug causes a protein to change shape.

描述（申请人提供）：生物医学的一个基本原理是小分子可以与特定受体结合以触发生理反应。我们试图了解当神经肌肉乙酰胆碱受体的神经递质结合位点在非传导和离子传导构象之间“门控”时发生的分子事件的能量本质。这些知识将帮助我们了解配体-蛋白质复合物的生物物理机制，并将提高我们为烟碱（和其他）受体设计新药的能力。基于结构的药物发现应考虑以下事实：药物作用取决于配体与受体的非活性构象和活性构象的差异结合。为了解决这一点，我们将使用单通道电生理学和动力学分析来研究未配体门控，这将使我们能够测量所有显着激活平衡常数，并确定两个递质结合位点中每个侧链的能量贡献。我们还将估计野生型和突变型受体中小配体探针的不同结合能。最终，这些知识将被用来设计对任意配体做出可预测反应的乙酰胆碱受体。公共健康相关性：该项目中概述的实验确立了工程蛋白质以可预测的方式对特定药物做出反应的基本原理。这些原理可应用于新药物的开发，以及了解药物导致蛋白质改变形状的机制。