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.

项目摘要：广泛影响：大电导钾通道（BK）等离子渠道是 负责可激发细胞的激发性质，例如神经元，骨骼肌细胞和心脏 肌肉细胞。对于BK通道，它们对膜电压和细胞内的双重敏感性 钙的变化使它们成为结合两种刺激以调节膜电压的理想工具。最近和 过去的工作表明，BK通道与对电压敏感的 - 经过钙的离子通道一起工作 用于细胞内信号。最近的工作还表明，BK通道中突变的患者可以 发展，神经和其他健康问题。因此，了解BK频道门控至关重要 重要性对于理解细胞内钙信号的调节方式。最近，有工作 这辩论了以前公认的电压传感器和钙之间独立的观念 传感器。在这里，该项目的目的是通过利用门中断的突变体来隔离每个辩论 栅极，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 电压传感器仅由于电压以及传感器之间的相互作用而没有贡献 将使用开放的孔，组成型通道。 AIM 2：组成性开放的渠道将用于 揭示由于开放通道而引起的完全调制电压传感器以及 传感器具有敞开孔的全部贡献。培训：该项目旨在教申请人如何 记录和分析门控电流。额外的培训将教申请人考虑投入输出 可能影响模型输出的建模和输入相互作用。该建议还将包括 与离子频道生物物理学专家互动的机会，以及分支为其他技术的分支 在离子通道生物物理学中，例如成像技术和分子动态模拟。旁边 彻底的专业发展方法，该提案将既有申请人需要的技能 成为一名贡献的职业科学家，并提高我们对离子门控机制的集体知识 频道。