STRUCTURE/FUNCTION ANALYSIS OF VOLTAGE-GATED CA CHANNELS

电压依赖性Ca ~（2+）通道控制钙离子进入细胞 生理和病理条件。他们是一个主要阶级的目标 高血压控制药物，它们是自身免疫性疾病的目标。 导致细胞死亡的反应，如可能发生在罕见疾病， 肌萎缩侧索硬化症以及更常见的疾病， I型糖尿病了解它们的多样性和复杂性 它们的功能至关重要。 这项研究的目的是将alpha 1的结构要素与 和Ca 2+通道β亚基的功能特性。的 使用的基本工具是通过重组DNA构建 突变体和嵌合分子的技术以及它们的测量 在非洲爪蟾卵母细胞中表达后相互作用和性质。 期望通过这些信息获得积极的信息 操作基于1。通过分子克隆获得 在我们的部分，两个同源的α 1 cDNA的，C型和E型， 区分电生理学和药理学特性， 能在非洲爪蟾卵母细胞中良好表达; 2.收购通过 我们的分子克隆了几个β亚基cDNA， 也在非洲爪蟾卵母细胞中表达，其中一些赋予 与α 1亚基共表达时， 非洲爪蟾卵母细胞;和3.通过PI的合作， Enrico Stefani博士，他将进行一次彻底而富有洞察力的 构建的电生理特性的分析 分子。 具体目标集中在通过构建嵌合体来阐明 alpha 1的主要结构-功能关系， 确定那些结构所需的钙失活的一些，但不是 其它的用于电压失活，用于通过磷酸化调节， 关键的药理学特性。 申请作为研究者发起的 研究项目补助金（IRPG）。伴侣补助金是史蒂芬尼博士喜欢的类型 II续期申请AR-38970。这些研究的结果可能 有助于设计更好的治疗药物。

Voltage grated Ca2+ channels control entry of Ca2+ into cells under physiologic and pathologic conditions. They are targets of a major class of hypertension controlling drugs, they are targets of autoimmune reactions that lead to cell death as may happen in rare diseases such as amyotrophic lateral sclerosis as well as in more frequent diseases such as type I diabetes. An understanding of their diversity, their complexity and their functioning is of central importance. The aim of the research is to relate structural elements of the alpha1 and beta subunits of Ca2+ channels to their functional properties. The basic tools to be used are the construction via recombinant DNA techniques of mutant and chimeric molecules and the measurement of their interaction and properties upon expression in Xenopus oocytes. The expectation that positive information will be obtained through these manipulations is based on 1. the acquisition through molecular cloning on our part of two homologous alpha1 cDNA's, type C and type E, that have distinguishing electrophysiological and pharmacological properties and can be expressed well in Xenopus oocytes; 2. the acquisition through molecular cloning on our part of several beta subunit cDNA's that can also be expressed in Xenopus oocytes and of which some impart distinguishing properties to the alpha1 subunits when co-expressed in Xenopus oocytes; and 3. the association through collaboration of the PI with Dr. Enrico Stefani, who will carry out a thorough and insightful analysis of the electrophysiological properties of the constructed molecules. Specific aims center on the elucidation through construction of chimeras of the major structure-function relations for alpha1 in an attempt to determine those structures required for Ca2+ inactivation of some but not others, for voltage inactivation, for regulation by phosphorylation and for key pharmacologic properties. The application is submitted as part of an Investigator-initiated Research Project Grant (IRPG). The companion grant is Dr. Stefani's type II renewal application AR-38970. Results from these studies may contribute to the design of better therapeutic agents.