Molecular mechanisms of Tandem Pore potassium channel gating and regulation

串联孔钾通道门控和调节的分子机制

• 批准号: 10798979
• 负责人: Paul Michael Riegelhaupt
• 金额: $ 15.17万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10798979
• 关键词: Address; Affect; Affinity; Architecture; Arrhythmia; Behavior; Binding Sites; Biological Assay; Biology; Biophysics; Blood Vessels; Cardiac; Cells; Combinatorics; Cryoelectron Microscopy; Crystallization; Cues; Data; Detergents; Development; Disease; Drug Targeting; Eicosanoids; Electrodes; Electrophysiology (science); Endocrine; Environment; Family; Goals; Human; Individual; Ion Channel; Ions; Knowledge; Lipid Binding; Lipids; Mass Spectrum Analysis; Measures; Mechanics; Membrane; Membrane Potentials; Mental Depression; Modeling; Molecular; Molecular Conformation; Movement; Mutate; Neurons; Nonesterified Fatty Acids; Output; Pharmaceutical Preparations; Pharmacology; Phospholipids; Physiological; Physiology; Play; Potassium; Potassium Channel; Regulation; Research; Resolution; Rest; Role; Signal Transduction; Site-Directed Mutagenesis; Specificity; Stretching; Structural Models; Structure; Temperature; Therapeutic; Therapeutic Agents; Transmembrane Domain; X-Ray Crystallography; chronic pain; chronic pain management; clinical practice; design; drug development; experimental study; improved; in vivo; mechanical force; mutant; mutation screening; nanodisk; novel; particle; patch clamp; potassium ion; protein reconstitution; proteoliposomes; response; small molecule; tool; voltage clamp

Abstract The goals of this project are to determine the molecular mechanisms that control activity of the tandem pore (K2P) family of potassium channels, with a focus on how K2Ps integrate a diverse set of incoming signals to regulate channel function. K2Ps are ion channels primarily responsible for producing background “leak” currents that set cellular resting membrane potential. Modulation of K2P activity directly affects cellular excitability and K2P channels have been implicated to play important roles in cardiac, neuronal, endocrine, and vascular biology. The mechanosensitive subfamily of K2Ps that are the focus of this proposal have been identified as potential drug development targets for treatment of cardiac arrhythmia, depression, and chronic pain, though efforts to develop small molecule modulators that target K2Ps have largely failed to produce high affinity and subtype selective agents. Meanwhile, endogenous lipids are known to modulate many K2P channels and show strong subtype specificity. The overall aim of this proposal is to examine the basic biology underlying K2P modulation by lipids and the related effects of membrane tension, with the long-term goal of using this knowledge to develop a framework for development of improved pharmacology against K2P channels. To achieve these goals, we will pursue a multifaceted approach that includes cryoEM structural studies, native mass spectrometry to define K2P/lipid interactions, and electrophysiological functional studies of K2P behavior. Our first aim will examine the mechanisms by which positive and negative allosteric membrane phosopholipids or free fatty acids are sensed by K2P channels, with a focus on the molecular details of lipid binding sites in the K2P structure. We will also address the basis for the interrelated impacts of allosteric lipids and membrane tension on K2P gating behavior. In aim2, we will explore the mechanism by which lipids, mechanosensitivity, and other K2P input signals control channel output at the ion conducting potassium selectivity filter, defining the molecular connectivity within the structural architecture of the K2P channel. Taken together, our studies will provide a detailed structural model of K2P gating and modulation that is broadly relevant to the basic biology of this important family of channels.

抽象的 该项目的目标是确定控制串联孔的分子机制 （k2p）钾渠道家族，重点是K2P如何将各种传入信号整合到 调节通道功能。 K2P是离子通道主要负责产生背景“泄漏”电流 那设定了细胞静止膜电位。 K2P活性的调节直接影响细胞令人兴奋和 K2P通道已被实施，以在心脏，神经元，内分泌和血管生物学中发挥重要作用。 K2P的机械敏感亚科是该提案的重点 药物开发目标用于治疗心律不齐，抑郁和慢性疼痛，尽管努力 开发针对K2P的小分子调节剂在很大程度上无法产生高亲和力和亚型 选择性代理。同时，已知内源性脂质可以调节许多K2P通道并显示强大的脂质 亚型特异性。该提案的总体目的是检查K2P调制的基本生物学 通过脂质和膜张力的相关作用，其长期目标是使用这些知识来发展 针对K2P渠道改善药理学的开发框架。为了实现这些目标，我们将 采用包括冷冻结构研究，天然质谱法的多面方法 K2P/脂质相互作用以及K2P行为的电生理功能研究。我们的第一个目标将研究 阳性和负变构磷脂或游离脂肪酸的机制 通过K2P通道，重点是K2P结构中脂质结合位点的分子细节。我们也会 解决变构脂质和膜张力对K2P门控行为的相互关联影响的基础。 在AIM2中，我们将探索脂质，机理敏感性和其他K2P输入信号控制的机制 导电钾选择性过滤器处的通道输出，定义了分子连通性 K2P频道的结构结构。综上所述，我们的研究将提供详细的结构模型 与这个重要渠道家族的基本生物学广泛相关的K2P门控和调节。