Structural basis of KATP Channel Gating

KATP通道门控的结构基础

• 批准号: 8131340
• 负责人: Show-Ling Shyng
• 金额: $ 30.8万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-06 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8131340
• 关键词: ATP-Binding Cassette Transporters; Address; Affect; Affinity; Basic Science; Binding; Cardiac; Cells; Chemicals; Childhood; Complex; Coupling; Data; Development; Diabetes Mellitus; Diagnosis; Diazoxide; Disease; Exhibits; Family; Fostering; Functional disorder; Genetic; Glucose; Goals; Health Sciences; Homeostasis; Human; Hyperinsulinism; Infant; Ion Channel; Kir6.2 channel; Knowledge; Lead; Link; Mediating; Membrane; Metabolic; Metabolism; MgADP; Molecular; Molecular Conformation; Mutagenesis; Mutate; Mutation; Myopathy; N-terminal; Neurologic; Neurons; Non-Insulin-Dependent Diabetes Mellitus; Nucleotides; Pancreas; Pathway interactions; Persistent Hyperinsulinemia Hypoglycemia of Infancy; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Physiological; Play; Potassium; Probability; Proteins; Publishing; Regulation; Research; Role; Scanning; Screening procedure; Signal Transduction; Signal Transduction Pathway; Structural Models; Structure-Activity Relationship; Sulfonylurea Compounds; Testing; Therapeutic Agents; Work; base; cell type; crosslink; human disease; innovation; insulin secretion; member; neonatal diabetes mellitus; novel; novel therapeutics; response; sulfonylurea receptor

DESCRIPTION (provided by applicant): ATP-sensitive potassium (KATP) channels couple cell metabolism to membrane excitability. They play a vital role in energy homeostasis by eliciting physiological response appropriate to metabolic signals. A member of the inwardly rectifying potassium (Kir) channel family, KATP channels are unique in requiring co-assembly of Kir6.2 with a sulfonylurea receptor (SUR), an ABC transporter, for functional expression. In pancreatic ¿- cells, KATP channels formed by Kir6.2 and SUR1 mediate glucose-stimulated insulin secretion. Mutations in Kir6.2 or SUR1 that reduce channel function cause congenital hyperinsulinism whereas those that increase channel function cause neonatal diabetes. Both Kir6.2 and SUR1 play an integral role in the channel's complex gating regulation. The long-term goal of this project is to understand the structure-function relationship of channel proteins with respect to gating. Work in the previous cycle has identified and elucidated the role of several structural features of the pore-forming subunit Kir6.2 in channel gating. In this renewal application, we will address the mechanisms of functional coupling between SUR1 and Kir6.2. SUR1 increases the open probability (Po) of Kir6.2, hypersensitizes Kir6.2 to the inhibitory effect of ATP and stimulatory effect of membrane phosphoinositides, and confers the effects of MgADP as well as the pharmacological agent sulfonylureas and diazoxide on Kir6.2. Functional coupling between SUR1 and Kir6.2 is thus essential to channel gating; yet significant knowledge gaps remain regarding the mechanisms by which SUR1 exerts its multiple effects on Kir6.2. The goal of this renewal application is to elucidate the mechanisms and structural basis that are responsible for functional coupling between SUR1 and Kir6.2. Based on our preliminary data, we propose a unifying hypothesis to explain how SUR1 imposes its multiple effects on Kir6.2 gating. Specifically, we hypothesize that SUR1 confers the intrinsic Po of KATP channels by stabilizing Kir6.2 in the PIP2-bound open state via molecular interactions between residues in the short cytoplasmic loops of TMD0 in SUR1 and those in the N-terminal domain of Kir6.2; and ATP, MgADP and pharmacological agents modulate channel activity in turn by changing the SUR1-Kir6.2 interface to strengthen or weaken channel-PIP2 interactions. We will interweave forward genetics approach employing disease mutations, guided mutagenesis screening, chemical cross-linking and structural modeling approaches to test the hypothesis. The research is innovative because it presents a novel concept to the field. The research is significant from both the human health and basic science standpoints. It will identify new disease mechanisms to directly facilitate diagnosis and treatment of several rare but devastating infant/childhood diseases and will lead to a better understanding of the structure-function relationship of the channel to foster new ideas on how to modulate channel activity to treat diseases caused by channel dysfunction, including type II diabetes. It will also lead to a better understanding of how a silent ABC transporter regulates an ion channel to advance both the ABC transporter and the ion channel fields. PUBLIC HEALTH RELEVANCE: The ATP-sensitive potassium (KATP) channels play a key role in linking metabolic signals to physiological responses in many cell types. Dysfunction of KATP channels causes human disease including diabetes, hyperinsulinism, cardiac myopathy and neurological deficits. The goal of this project is to understand the molecular basis underlying the ability of the channel to open or close according to metabolic signals to facilitate development of novel therapeutic agents for disease caused by channel dysfunction.

描述（由申请方提供）：ATP敏感性钾（KATP）通道将细胞代谢与膜兴奋性偶联。它们通过引发与代谢信号相适应的生理反应，在能量稳态中起着至关重要的作用。作为内向整流钾（Kir）通道家族的一员，KATP通道在需要Kir6.2与磺酰脲受体（SUR）（ABC转运蛋白）共组装以实现功能表达方面是独特的。在胰腺细胞中，由Kir6.2和SUR 1形成的KATP通道介导葡萄糖刺激的胰岛素分泌。Kir6.2或SUR 1突变降低通道功能导致先天性高胰岛素血症，而增加通道功能导致新生儿糖尿病。Kir6.2和SUR 1在通道的复杂门控调节中起着不可或缺的作用。这个项目的长期目标是了解通道蛋白与门控的结构-功能关系。上一个周期的工作已经确定并阐明了成孔亚基Kir6.2的几个结构特征在通道门控中的作用。在这次更新申请中，我们将讨论SUR 1和Kir6.2之间的功能耦合机制。SUR 1增加Kir6.2的开放概率（Po），使Kir6.2对ATP的抑制作用和膜磷酸肌醇的刺激作用超敏，并赋予MgADP以及药理学试剂磺酰脲类和二氮嗪对Kir6.2的作用。因此，SUR 1和Kir6.2之间的功能耦合对于通道门控是必不可少的;然而，关于SUR 1对Kir6.2发挥其多重作用的机制，仍然存在重大的知识空白。该更新申请的目标是阐明负责SUR 1和Kir6.2之间功能偶联的机制和结构基础。基于我们的初步数据，我们提出了一个统一的假设来解释SUR 1如何对Kir6.2门控产生多重影响。具体地说，我们假设SUR 1通过SUR 1中TMD 0的短胞质环中的残基与Kir6.2的N-末端结构域中的残基之间的分子相互作用，将Kir6.2稳定在PIP 2结合的开放状态，从而赋予KATP通道的内在Po;和ATP，MgADP和药理学试剂通过改变SUR 1-Kir6.2界面以加强或减弱通道-PIP 2相互作用来调节通道活性。我们将交织正向遗传学方法采用疾病突变，诱变筛选，化学交联和结构建模方法来验证假设。这项研究是创新的，因为它提出了一个新的概念，该领域。从人类健康和基础科学的角度来看，这项研究都具有重要意义。它将确定新的疾病机制，以直接促进几种罕见但毁灭性的婴儿/儿童疾病的诊断和治疗，并将导致更好地了解通道的结构-功能关系，以促进关于如何调节通道活性以治疗由通道功能障碍引起的疾病（包括II型糖尿病）的新思路。这也将导致更好地了解沉默的ABC转运蛋白如何调节离子通道，以推进ABC转运蛋白和离子通道领域。 公共卫生相关性：ATP敏感性钾通道（KATP）在许多细胞类型中将代谢信号与生理反应联系起来起着关键作用。KATP通道的功能障碍导致人类疾病，包括糖尿病、高胰岛素血症、心肌病和神经缺陷。该项目的目标是了解通道根据代谢信号打开或关闭能力的分子基础，以促进开发用于通道功能障碍引起的疾病的新型治疗药物。