To Clarify the Debates Surrounding BK Gating

项目摘要：影响广泛：离子通道，如大电导钾通道(BK) 负责兴奋性细胞的兴奋性，如神经元、骨骼肌细胞和心脏 肌肉细胞。特别是BK通道，其对膜电压和细胞内电压的双重敏感性 钙离子的变化使它们成为结合两种刺激来调节膜电压的完美工具。最近和 过去的工作表明，BK通道与电压敏感的、可调节的钙离子通道一起工作 用于细胞内信号传递。最近的研究还表明，BK通道突变的患者可以 发育、神经和其他健康问题。因此，了解BK通道门控是非常重要的 了解细胞内钙信号是如何被调节的很重要。最近，有一些工作 它辩论了以前被接受的电压传感器和钙之间相互独立的想法- 传感器。在这里，这个项目的目的是通过利用门中断的突变体来分离每个 栅极、电压传感器和钙传感器之间的相互作用。目标1：揭示未调制的 仅由电压引起的电压传感器以及传感器之间的相互作用，而不受 将使用开孔、结构闭合的通道。目标2：将利用结构性开放的渠道 揭示了由于开放的通道和调制之间的相互作用而产生的完全调制的电压传感器 传感器的全部贡献来自一个开放的气孔。培训：该项目旨在教申请者如何 记录和分析门控电流。额外的培训将教会申请人思考投入-产出 可能影响模型输出的建模和输入交互作用。这项提案还将包括 有机会与离子通道生物物理学方面的专家互动，以及学习使用的其他技术 在离子通道生物物理学方面，如成像技术和分子动力学模拟。旁边是一个 通过全面的职业发展途径，这项建议将既培养申请人所需的技能 成为一名有贡献的职业科学家，同时增加我们对离子门控机制的集体知识 频道。

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.