To Clarify the Debates Surrounding BK Gating

澄清围绕 BK 门控的争论

• 批准号: 10450659
• 负责人: Frank Yeh
• 金额: $ 4.35万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10450659
• 关键词: Action Potentials; Affinity; Binding; Binding Sites; Biophysics; Calcium; Calcium Signaling; Cardiac Myocytes; Cell membrane; Cells; Child; Coupled; Coupling; Cryoelectron Microscopy; Development; Freedom; Fright; Health; Imaging Techniques; Impairment; Intracellular Membranes; Investigation; Ion Channel; Knowledge; Long-Term Depression; Long-Term Potentiation; Membrane; Modeling; Molecular; Molecular Conformation; Movement; Muscle Fibers; Mutation; Nature; Nervous System Physiology; Neurologic; Neurons; Output; Patients; Potassium Channel; Process; Proteins; Scientist; Seizures; Shapes; Signal Transduction; Site; Stimulus; Structure; Techniques; Training; Work; career; developmental disease; experimental study; large-conductance calcium-activated potassium channels; mutant; protein structure; response; sensor; simulation; skills; tool; training project; vesicular release; voltage

Project Summary: Broad Impact: Ion channels such as the big conductance potassium channels (BK) are responsible for the excitable nature of excitable cells, such as neurons, skeletal muscle cells, and cardiac muscle cells. For BK channels specifically, their dual sensitivity to both membrane voltage and intracellular calcium changes make them a perfect tool to combine both stimuli to modulate membrane voltage. Recent and past work show that BK channels work alongside voltage-sensitive, -modulated ion channels that pass calcium for intracellular signaling. Recent work also shows that patients with mutations in BK channels can have developmental, neurological, and other health issues. As such, understanding BK channel gating is of utmost importance to understanding how intracellular calcium signaling is modulated. Recently, there are have work that debates the formerly accepted idea of independence between the voltage-sensor and the calcium- sensors. Here, this project will aim to satisfy this debate by utilizing gate-disrupted mutants to isolate each interaction between the gate, the voltage-sensor, and the calcium-sensor. Aim 1: To reveal the unmodulated voltage-sensor due to only voltage and the interactions between the sensors without contributions from an open pore, constitutively-closed channels will be used. Aim 2: Constitutively-open channels will be used to reveal the fully modulated voltage-sensor due to an open channel and the modulated interactions between the sensors with full contributions from an open pore. Training: This project aims to teach the applicant how to record and analyze gating currents. Additional training will to teach the applicant to think about input-output modelling and input interactions that might influence output of a model. This proposal will also include opportunities to interact with experts in ion channel biophysics as well as to branch into other techniques used in ion channel biophysics such as imaging techniques and molecular dynamic simulations. Alongside a thorough approach to professional development, this proposal will both develop skills needed for the applicant to be a contributing career scientist as well as increase our collective knowledge on gating mechanisms of ion channels.

项目概述：广泛影响：离子通道，如大电导钾通道（BK）， 负责可兴奋细胞（例如神经元、骨骼肌细胞和心脏细胞）的兴奋性 肌肉细胞对于BK通道，它们对膜电压和细胞内电压的双重敏感性， 钙的变化使它们成为联合收割机结合两种刺激来调节膜电压的完美工具。最近和 过去的研究表明BK通道与电压敏感的、可调节的离子通道一起工作， 用于细胞内信号传导。最近的研究还表明，BK通道突变的患者可以有 发育、神经和其他健康问题。因此，了解BK通道门控是至关重要的 重要的是了解细胞内钙信号是如何调制。最近有工作 这与以前接受的电压传感器和钙离子之间独立的观点存在争议， 传感器.在这里，这个项目的目标是通过利用门中断突变体来分离每个 门、电压传感器和钙传感器之间的相互作用。目的1：揭示未调制的 电压传感器由于只有电压和传感器之间的相互作用，而没有来自 将使用开孔、组成封闭的通道。目标2：将利用宪法规定的公开渠道， 揭示了完全调制的电压传感器，由于开放的通道和调制的相互作用之间的 传感器具有来自开孔的全部贡献。培训：本项目旨在教申请人如何 记录并分析门控电流。额外的培训将教申请人思考投入产出 可能影响模型输出的建模和输入交互。该提案还将包括 有机会与离子通道生物物理学专家进行互动，以及分支到其他技术使用 离子通道生物物理学，如成像技术和分子动力学模拟。并配以香味四溢 通过对专业发展的深入研究，该提案将培养申请人所需的技能 成为一名有贡献的职业科学家，并增加我们对离子门控机制的集体知识 渠道