Development /use of peptidic inhibitors for Kir channels

Kir 通道肽抑制剂的开发/使用

• 批准号: 6727515
• 负责人: ZHE LU
• 金额: $ 27.58万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6727515
• 关键词: bacteriophage M13; cell membrane; chemical synthesis; inhibitor /antagonist; membrane proteins; nucleic acid probes; peptide library; phage display; potassium channel; protein binding; protein structure function; toxicant interaction; toxin

DESCRIPTION: The long term goal of our research is to develop effective inhibitors for inward-rectifier K+ channels and use them as molecular probes to explore the molecular mechanisms underlying the function of these ion channels. We have recently identified tertiapin, a honeybee toxin, as a nanomolar-affinity protein-inhibitor for some inward-rectifier K+ channels. Tertiapin, whose structure is known, inhibits the channels by binding to the external vestibule of the K+ conduction pore. In Aim 1A, we will first use tertiapin as a probe to explore the molecular architecture of the outer pore in inward-rectifier K+ channels. By identifying the pairing relation between channel and toxin residues with thermodynamic mutant cycle analysis, we will be able to assign each channel residue pairing with a toxin residue to a location in space with respect to the toxin structure, and thus to delineate the architecture of the outer pore. We will then in Aim 1B examine the mechanism underlying the specificity of channel-toxin interactions. With this knowledge we will design inhibitors with higher channel specificity (Aim 2A), and will also employ phage display, a type of very powerful combinatorial peptide technology, to select specific channel-inhibiting peptides from random peptide libraries (Aim 2B). The proposed fundamental studies also have important medical implications. Inward-rectifier K+ channels are significant both physiologically and pathophysiologically and represent important pharmacological targets. The inhibitors that we develop will be powerful tools both for investigating the physiology and/or pathophysiology of the channels and for developing drugs that specifically target these channels.

描述：我们研究的长期目标是发展有效 内向置换剂K+通道的抑制剂，并将其用作分子探针 探索这些离子通道功能的基础的分子机制。 我们最近已经确定蜜蜂毒素的tertiapin是一个 某些内部旋转剂K+通道的纳摩尔 - 亲密蛋白抑制剂。 Tertiapin的结构已知，通过与 K+传导孔的外侧前庭。在AIM 1A中，我们将首先使用 tertiapin作为探索外孔的分子结构的探针 内向置换剂K+通道。通过确定配对之间的关​​系 带有热力学突变循环分析的通道和毒素残基，我们将是 能够将每个通道残基与毒素残留物分配到一个位置 在太空相对于毒素结构，从而描绘 外孔的建筑。然后，我们将在AIM 1B检查机制 基础通道毒素相互作用的特异性。有了这些知识 我们将设计具有较高频道特异性（AIM 2A）的抑制剂，并将 还采用噬菌体显示，一种非常强大的组合肽 技术，从随机肽中选择特定的通道抑制肽 图书馆（AIM 2B）。 拟议的基础研究也具有重要的医学意义。 内向促进剂K+通道在生理和 病理生理，代表重要的药理靶标。这 我们开发的抑制剂将是强大的工具 渠道的生理和/或病理生理学以及开发药物 专门针对这些渠道。