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：
10537846
负责人：
Elizabeth Dione Kim
金额：
$ 6.76万
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/10537846
关键词：
Affinity Agonist Algorithms Binding Binding Sites Bioinformatics Biological Models Cells Communication Complex Coupled Coupling Cryoelectron Microscopy Cues Cyclic Nucleotides Data Defect Development Disease Doctor of Philosophy Drug Binding Site Drug Design Drug Targeting Electrodes 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 Propofol Protein Family Protein Isoforms Proteins Regulation Role Route Sequence Alignment Severity of illness Signal Transduction Specificity Structure Structure-Activity Relationship Testing Validation analog antagonist base density design drug discovery experimental study 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.

项目概要环核苷酸调节的离子通道是一类在许多生理学中具有重要作用的蛋白质。过程，包括心脏调节、神经信号传导和疼痛感知。新的发现由于多种原因，针对不同离子通道的药物是众所周知的困难，因为重点是传统的正位激动剂和拮抗剂一直占主导地位。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。检测能力并分离变构相互作用网络的功能可以在设计中提供更有针对性的方法用于环核苷酸调节通道的变构药物。在第一个目标中，我将识别变构并对其进行分类使用协同进化分析的网络。诱变和定量电生理学测量将用于探测不同残基位置如何有助于通道门控中必要的能量耦合。下一个，我们将使用这个独特的信息来获得新的通道过渡状态。这一目标的总体目标是通过实验定义功能调节环核苷酸调节离子通道的变构网络验证这些网络的功能，并利用这些信息来获得难以解析的构象状态。在第二个目标中，我将寻求发现已知的环化变构调节剂的未知结合位点。核苷酸调节的离子通道。使用冷冻电镜，我们将确定复杂的通道结构建立了变构调节剂并定义了它们与目标 1 中的变构网络的相互作用。我们将使用诱变和两个电极电压钳验证结合位点。该目标将证明可行性针对变构网络的药物设计策略。