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Probing the structure and function of the intracellular domain of cys-loop receptors

探讨cys环受体胞内结构域的结构和功能

基本信息

批准号：
10540405
负责人：
Michaela Jansen
金额：
$ 38.25万
依托单位：
TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-15 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10540405
关键词：
Address Alzheimer&apos s Disease Amino Acids Anesthetics Anti-Anxiety Agents Antiemetics Antiepileptic Agents Anxiety Anxiety Disorders Architecture Atherosclerosis Attention Binding Biochemical Biological Assay Brain Cations Cause of Death Cell Line Cell membrane Characteristics Chimera organism Choline Cholinergic Receptors Clinical Coupled Cryoelectron Microscopy Dementia Diabetes Mellitus Disease Drug Design Drug Targeting Electrophysiology (science)Environment Epilepsy Eukaryota Extracellular Domain Family Family member Foundations Funding Future Glycine Heart Diseases Homo Human Immobilization Inflammatory Inflammatory Bowel Diseases Investigation Ion Channel Ion Channel Gating Knowledge Length Ligands Link Mediating Membrane Transport Proteins Mental Depression Methods Modification Molecular Molecular Chaperones Molecular Conformation Muscle relaxants Myasthenia Gravis Neurodegenerative Disorders Neurologic Neurotransmitter Receptor Nicotine Dependence Pain Parkinson Disease Peptides Pharmaceutical Preparations Plant Resins Proteins Publishing Resistance Role Schizophrenia Sepsis Serotonin Shapes Sodium Chloride Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Structure Surface Tertiary Protein Structure Testing Tissues Transmembrane Domain Western Blotting alpha helix burden of illness clinical effect conformational conversion design disability esterase experimental study extracellular gamma-Aminobutyric Acid inhibitor innovation insight maltose-binding protein nervous system disorder neuropsychiatric disorder novel overexpression pharmacologic prevent protein protein interaction protein purification receptor smoking cessation targeted treatment therapeutic development therapy design

项目摘要

Project Summary Pentameric ligand-gated ion channels (pLGICs), also called Cys-loop receptors in eukaryotes, include the receptors for acetylcholine, serotonin, GABA and glycine. They are involved in numerous neuropsychiatric, neurologic, and inflammatory diseases. All Cys-loop receptor family members in metazoans contain three structural domains: an extracellular domain (ECD), a transmembrane domain (TMD), and an intracellular domain (ICD). The ECD and TMD are architecturally conserved between receptor families. The ICD is poorly conserved in both length and amino-acid composition and, for many subunits, contains large regions of predicted structural disorder. Due to the challenge of working with the ICDs of pentameric receptors, they have been essentially overlooked in terms of rigorous mechanistic characterization and direct exploration as pharmacological targets. The ICD is involved in finetuning plasma membrane expression levels, targeting, and function, in part mediated by protein-protein interactions (PPI) for example with chaperones. We envision that mechanistic knowledge of chaperone-mediated modulation will uncover new PPI drug targets. Through our published studies of the ICD we defined a linker that allowed for structure determination of the first in class structures of homo- and heteropentameric GABAA receptors. We also established that the ICD alone assembles into pentamers, establishing a novel role for the ICD in oligomeric assembly. Here, we propose a multi-layered approach leveraging both soluble ICD chimeras and full-length receptors together with results obtained during the previous funding period in careful consideration of recently-published pLGIC structures. We discovered that the resistance to inhibitors of choline esterase (Ric-3) chaperone binds to a 24-amino acid L1-MX segment of 5-HT3A subunits. With the new proposed specific aims (SA) we will: (SA1) determine the role of the L1-MX segment in RIC-3 modulation of cationic pLGIC assembly, (SA2) identify novel protein-protein interactions mediated via cationic ICDs, and (SA3) characterize the mechanism by which opening of cation-conducting pLGICs involves translocation of the L1-MX segment through the MA-helix framed portals. In each aim, we will use biochemical and electrophysiological methods, coupled with overexpressed and purified proteins or proteins in their cellular environment. We anticipate that our studies will provide the basis for a novel class of targets for therapeutic development, PPI modulators for pentameric channel intracellular domains.

项目总结