CHARACTERIZATION OF ANTIARRHYTHMIC DRUG BINDING SITES IN VOLTAGE GATED CHANNELS

电压门控通道中抗心律失常药物结合位点的表征

• 批准号: 6242338
• 负责人: DAVID T YUE
• 金额: $ 24.57万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6242338
• 关键词: activation analysis; aminoacid; antiarrhythmic agent; drug design /synthesis /production; electrophysiology; genetic manipulation; potassium channel; site directed mutagenesis; sodium channel; transfection; voltage /patch clamp; voltage gated channel

Voltage-gated channels are not only essential to countless physiological functions in heart, but also serve as the molecular target of diverse anti-arrhythmic agents. With the recent cloning, sequencing, and expression of voltage-gated Na and K channels, the stage is now set to identify the key amino acid residues primarily involved in anti- arrhythmic action on voltage-gated Na and K channels. In particular, wild-type and mutant versions of both a voltage-gated Na channel derived from adult skeletal muscle (mu1, Trimmer et al., 1990), as well as a voltage-gated K channel derived from human heart (HK1, or Kv1.4, Tamkun et al., 1991) will be expressed in a mammalian cell line and probed by patch-clamp methods to detect functional changes in the effects of internally applied QX-314 (for mu1) and clofilium (for HK1), permanently charged versions of anti-arrhythmic agents. Mutations will be made according to recent work localizing the probable site of action of internally active anti-arrhythmic agents to the cytoplasmic vestibule of Na and K channels (Gingrich et al., 1993; Backx et al., 1992, Choi et al., 1993). Unmasking the arrangement of amino acids that are important for drug action would be a crucial prerequisite to unravelling the mechanism of anti-arrhythmic action at the molecular level. Such understanding would be invaluable in rational drug therapy and design.

电压门控通道不仅对无数生理必不可少 在心脏中发挥作用，但也充当多种多样的分子靶标 抗心律失常剂。 随着最近的克隆，测序和 电压门控的Na和K通道的表达，现在设置为 确定主要涉及抗抗的关键氨基酸残基 对电压门控的Na和K通道的心律不齐。 尤其， 衍生的电压门控通道的野生型和突变版本 从成年骨骼肌（Mu1，Trimmer等，1990）以及 电压门控的K通道源自人心（HK1或KV1.4，Tamkun） 等，1991）将在哺乳动物细胞系中表达，并通过 贴片钳方法检测功能变化的影响 内部应用QX-314（用于MU1）和氯富尔（用于HK1），永久 抗心律失常剂的充电版本。 突变会发生 根据最近的工作，将可能的行动站点定位 内部活跃的抗心律失常剂 Na和K通道（Gingrich等，1993; Backx等，1992； Choi et Al。，1993）。 揭示重要的氨基酸的排列 对于毒品作用将是揭露毒品的至关重要的先决条件 抗心律失常作用在分子水平上的机制。 这样的 理解在理性的药物治疗和设计中是无价的。