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The molecular architecture and mechanism of the Proton Activated Chloride (PAC) Channel.

质子活化氯化物 (PAC) 通道的分子结构和机制。

基本信息

批准号：
10157439
负责人：
Makayla Freitas
金额：
$ 4.35万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-28 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10157439
关键词：
ASIC channel Acidosis Acids Address Amino Acids Anions Architecture Biochemical Biological Assay Biological Process Brain C-terminal Cell Death Cells Cerebral Ischemia Characteristics Chloride Channels Chloride Ion Chlorides Code Cognitive Cryoelectron Microscopy Development Drug Targeting Electrophysiology (science)Environment Event Family Foundations Future Genes Goals Home environment Homeostasis Institutes Investigation Ion Channel Ion Channel Gating Ions Ischemia Ischemic Brain Injury Ischemic Stroke Knock-out Knowledge Ligands Link Mediating Membrane Proteins Mentorship Modeling Molecular Mutagenesis Mutate Mutation N-terminal Nervous system structure Neuraxis Neuronal Injury Neurons Neurosciences Pacific Northwest Physiological Physiological Processes Physiology Play Prevention Property Protein Biochemistry Protons Resolution Rest Role Scientist Senior Scientist Shapes Sodium Stroke Structure Structure-Activity Relationship Swelling Therapeutic Agents Tissues Translating Traumatic Brain Injury alpha helix base biophysical properties biophysical tools brain tissue career cell growth regulation chemical property desensitization design experimental study extracellular graduate student human tissue insight knowledge base millisecond mouse model mutant neuron loss novel novel therapeutics particle physical property protein folding response skills stoichiometry stroke survivor structural biology structured data therapeutic development therapeutic target

项目摘要

Project Summary Ischemic strokes can cause long-term cognitive damage, leading to reduced mobility in nearly half of stroke survivors. Brain tissue damage that occurs during and preceding an ischemic event is often largely in part due to severe local tissue acidosis. While the molecular mechanism of how acidosis leads to tissue damage is largely unknown, proton-gated ion channels are thought to play a role. A novel proton-gated chloride channel has been recently identified as the previously uncharacterized gene, TMEM206, now commonly referred to as the Proton-Activated Chloride (PAC) Channel. While recent studies have implicated PAC in acid-induced cell death, there exists no molecular justification of the channel’s proton-activated chloride currents. This proposal will integrate electrophysiological, biochemical, and high-resolution structural experiments to elucidate the structure-based mechanisms that govern the function of PAC. In support of this goal, I will first obtain the high- resolution structures of PAC’s resting and active functional state using single particle cryo-electron microscopy (Cryo-EM). These structures will provide fundamental insights into the architecture, stoichiometry, and unique protein folding of PAC. This information will also expand our knowledge surrounding the physical and chemical properties of proton-gated ion channels, as well as broadly inform the structure/function relationship of ion channels. I will then ascertain the molecular underpinnings of PAC’s pH-dependent mechanism and pore properties by probing PAC’s function using structure-directed mutagenesis and electrophysiological experiments. These experiments will establish a link between the molecular architecture and physiology of PAC. Ultimately, this proposal will define structure-based, biochemical mechanisms for PAC’s function, which will lay the foundation for future studies and may inform the development of therapeutic agents to mitigate neuronal damage in ischemic events. As a neuroscience graduate student whose goal is to become an independent academic scientist that will study the structure/function of ligand-gated ion channels of the nervous system, this project will directly expand my knowledge base and technical skillset in ion channels, membrane protein biochemistry, electrophysiology, and cryo-EM. The study into the molecular architecture and mechanism of PAC will be pursued under the mentorship of Dr. Eric Gouaux, an expert in ligand-gated ion channels and leader in membrane protein structural biology. Dr. Gouaux is a senior scientist at the Vollum Institute at OHSU, an electrophysiology powerhouse that is home to one of three national centers for Cryo-EM, Pacific Northwest Center for Cryo-EM (PNCC). Taken together, the project will not only illuminate critical insights into a novel proton-activated channel, but will also provide me with the necessary knowledge, biophysical tool-kit, and professional skills I need to achieve my long-term career goal.

项目总结