Identification of potential drug binding sites within allosteric networks in cyclic nucleotide modulated channels

环核苷酸调节通道变构网络内潜在药物结合位点的鉴定

基本信息

批准号：
10704557
负责人：
Elizabeth Dione Kim
金额：
$ 7.18万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-24 至 2024-08-23
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10704557
关键词：
Affinity Agonist Algorithms Binding Binding Sites Bioinformatics Biological Models Cells Classification Communication Complex Coupled Coupling Cryoelectron Microscopy Cues Cyclic Nucleotides Data Defect Development Disease Doctor of Philosophy Drug Binding Site Drug Design Drug Targeting Electrodes Electronic Medical Records and Genomics Network Electrophysiology (science)Equilibrium Evaluation Exposure to FDA approved Family Frequencies Functional disorder Future Genomics Goals HCN1 channel HCN1 gene Heart Human Genome Human body Hyperactivity Individual Ion Channel Link Lipids Location Measurement Measures Methods Molecular Conformation Mutagenesis Mutate Mutation Pathway Analysis Pharmaceutical Preparations Physiological Processes Population Positioning Attribute Postdoctoral Fellow Propofol Protein Family Protein Isoforms Proteins Reaction Regulation Role Sequence Alignment Severity of illness Signal Transduction Specificity Structure Structure-Activity Relationship Testing Validation analog antagonist design drug discovery experimental study genomic predictors inhibitor interest member mutant nanodisk neurotransmission new therapeutic target novel pain perception particle patch clamp practical application receptor reconstitution side effect small molecule structural biology voltage voltage clamp

项目摘要

PROJECT SUMMARY Cyclic nucleotide modulated ion channels are a class of proteins that have important roles in many physiological processes, including regulation of the heart, neuronal signaling, and pain perception. The discovery of new drugs targeting different ion channels is notoriously difficult for a host of reasons, as the focus on traditional orthosteric agonists and antagonists has been dominant. Overall, this proposal unifies genomic, functional, and structural methods to reveal how specific allosteric interactions govern mechanistic function. The ability to detect and isolate the function of networks of allosteric interactions can provide a more focused approach in the design of allosteric drugs for cyclic nucleotide modulated channels. In the first aim, I will identify and classify allosteric networks using coevolution analysis. Mutagenesis and quantitative electrophysiology measurements will be used to probe how different residue positions contribute to requisite energetic coupling in channel gating. Next, we will use this unique information to obtain novel channel transition states. The overall goal of this aim is to define allosteric networks that functionally regulate cyclic nucleotide modulated ion channels, experimentally validate these networks functionally, and use this information to obtain difficult-to-resolve conformational states. In the second aim, I will seek to uncover unknown binding sites for known allosteric modulators of cyclic nucleotide modulated ion channels. Using cryoEM, we will determine the structure of channels in complex with established allosteric modulators and define their interaction with the allosteric networks in aim 1. We will validate the binding site using mutagenesis and two electrode voltage clamp. This aim will demonstrate feasibility of drug design strategies targeting allosteric networks.

项目摘要环状核苷酸调制离子通道是一类蛋白质，在许多生理学中具有重要作用过程，包括心脏的调节，神经元信号传导和疼痛感知。发现新的众所周知，针对不同离子渠道的药物出于多种原因很难，因为关注传统正常的激动剂和对手一直是主导的。总体而言，该提案统一了基因组，功能和揭示特定变构相互作用如何控制机械功能的结构方法。检测能力并隔离变构相互作用网络的功能可以在设计中提供更集中的方法循环核苷酸调制通道的变构药物。在第一个目标中，我将识别和分类变构使用协同进化分析的网络。诱变和定量电生理测量将是用于探测不同的残基位置如何在通道门控中有必要的能量耦合。下一个，我们将使用这些独特的信息来获得新颖的渠道过渡状态。这个目标的总体目标是定义在功能调节环状核苷酸调制离子通道的变构网络，实验在功能上验证这些网络，并使用此信息获得难以解决的构象状态。在第二个目标中，我将寻求发现已知循环变构调节剂的未知绑定位点核苷酸调制离子通道。使用Cryoem，我们将确定通道的结构建立的变构调节器并在AIM 1中定义了与变构网络的互动。我们将使用诱变和两个电极电压夹验证结合位点。这个目标将证明可行性针对变构网络的药物设计策略。