FUNCTIONAL DOMAINS OF POTASSIUM CHANNEL SUBUNIT PROTEINS

钾通道亚基蛋白的功能域

• 批准号: 2839356
• 负责人: Paul Pfaffinger
• 金额: $ 27.74万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-05-01 至 2000-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2839356
• 关键词: Xenopus; Xenopus oocyte; gene expression; laboratory rabbit; membrane structure; neurons; potassium channel; protein biosynthesis; protein structure function; site directed mutagenesis; voltage gated channel

DESCRIPTION: K+ channels are an important target for drugs to treat a variety of disorders, including high blood pressure, diabetes, asthma, multiple scleroisis, and others. In addition, the genetic disease, Periodic Ataxia and certain forms of Long Q-T syndrome, have been linked to specific point mutations in K+ channel proteins. Despite the importance these channels have in maintaining normal physiology, and their net therapeutic importance, relatively little is known about how K+ channel assembly by the N-terminal T1 domain. The experiments the investigator will perform in this proposal are motivated by the progress he has achieved in understanding the assembly mechanisms of voltage gated K channels from the original proposal. In this new project, the investigator will determine the crystallographic structure of the T1 domain protein in collaboration with Dr. Senyon Choe, confirm the structural model in isolated T1 domain proteins and full length channels, identify critical residues and interactions for K channel assembly, and characterize the alterations in channel assembly identity produced by T1 domain mutations. The long term goal is to understand how information needed to encode the precise electrophysiological properties of the nervous system are encoded by structural differences in the T1 domain of K channel subunit proteins. This goal will be addressed by experiments that will satisfy the following Specific Aims: 1) Determination of the structure of the tetrameric T1 domain and identification of functionally significant regions of the T1 domain protein; 2) analysis of residues that are functionally important for T1 domain tetramerization and confirmation of the T1 domain structure; and 3) analysis of the T1 domain in the full length channel subunit protein and its role in controlling channel protein assembly. The general approach in these studies is to integrate structural information with functional information on wild type and mutant T1 domain assembly in isolated protein preparations, and for full length subunit protein assembly, and intact cells. By careful characterization of the changes in assembly properties caused by these individual mutations, the investigators can dissect the functional roles for specific residues and residue interactions in producing the ordered assembly of K channel subunit protein in the nervous system.

描述：K+通道是药物治疗糖尿病的重要靶点。 各种疾病，包括高血压，糖尿病，哮喘， 多发性硬化症等。 此外，遗传性疾病，周期性 共济失调和某些形式的长Q-T综合征，已被链接到特定的 K+通道蛋白的点突变。 尽管这些重要性 经络在维持正常生理功能方面的作用， 重要的是，相对较少的是知道如何K+通道组装由 N-末端T1结构域。 研究人员将在此进行的实验 他的提议是由他在理解 组装机制的电压门控K通道从原来的建议。 在这个新项目中，研究人员将确定晶体学 与Senyon Choe博士合作， 证实了分离的T1结构域蛋白和全长的结构模型 通道，确定K通道的关键残基和相互作用 组件，并表征通道组件标识的更改 由T1结构域突变产生。 长期目标是了解 编码精确的电生理特性所需的信息 神经系统由T1结构域的结构差异编码， K通道亚单位蛋白质类。 这一目标将通过实验来实现， 将满足以下具体目的：1）确定结构 的四聚体T1结构域和鉴定的功能显着 T1结构域蛋白的区域; 2）分析 对于T1结构域四聚化和确认 T1结构域结构;和3）全长T1结构域的分析 通道亚基蛋白及其在控制通道蛋白中的作用 组装件. 这些研究的一般方法是整合结构 关于野生型和突变T1结构域功能信息 在分离的蛋白质制备物中的组装，以及对于全长亚基 蛋白质组装和完整细胞。 通过仔细描述 由这些单独的突变引起的组装属性的变化， 研究人员可以剖析特定残基的功能作用， K通道亚基有序组装中的残基相互作用 神经系统中的蛋白质。