STRUCTURE/FUNCTION OF ALPHA1 ADRENERGIC RECEPTORS

ALPHA1 肾上腺素能受体的结构/功能

• 批准号: 2271071
• 负责人: Kenneth P Minneman
• 金额: $ 20.38万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2271071
• 关键词: alpha adrenergic receptor; antisense nucleic acid; calcium flux; complementary DNA; cyclic AMP; enzyme activity; genetic library; molecular cloning; neuropharmacology; nucleic acid sequence; phospholipase A2; phospholipase D; protein structure function; receptor coupling; transfection; transfection /expression vector

alpha1-Adrenergic receptors (ARs) play important roles in control of blood pressure, nasal congestion, muscle growth, and other processes. Two native subtypes (alpha1A and alpha1B) can be distinguished pharmacologically, and three subtypes have been cloned (alpha1B, alpha1C, and alpha1D). Although one of the cloned subtypes (alpha1D) was originally thought to encode the pharmacologically defined alpha1A subtype, recent data suggests that this is unlikely. The relationship between the cloned receptors and the native subtypes must be understood, and any additional cDNA clones obtained, before the biochemical and functional roles of alpha1-AR subtypes can be clearly understood. Human SKNMC neuroepithelioma cells express at least two alpha1-ARs, resembling the alpha1A, and alpha1B subtypes. We have isolated, expressed, and partially characterized a full length chimeric genomic/cDNA clone from this cell line which is homologous to the rat alpha1D. We will use sense expression and antisense knockout of this and other human clones in SKNMC cells to clarify the relationship of the known clones to the pharmacological alpha1A subtype. If they are unrelated, as expected, we will obtain a full length alpha1A cDNA by expression cloning. With these clones we will be able to clearly define the drug specificities, signalling mechanisms, and interactions of alpha1-AR subtypes. Our major goals include: 1. To clearly define the relationship between the 3 known clones and the pharmacologically defined alpha1A subtype; 2. To obtain a full-length cDNA for the alpha1A subtype by expression cloning; 3. To determine whether different alpha1-AR receptor subtypes preferentially couple to different second messenger systems; and 4. To determine whether different subtypes co-existing in the same cell have additive, synergistic or redundant interactions which may contribute to the confusion about the number and properties of alpha1-AR subtypes. These experiments will clarify the number, signalling mechanisms, and interactions of alpha1-AR subtypes. Since drugs acting on these receptors have considerable therapeutic importance, such information may have direct clinical impact.

α 1-肾上腺素能受体（AR）在控制 血压，鼻塞，肌肉生长和其他过程。 可以区分两种天然亚型（α 1A和α 1B） 已经克隆了三种亚型（α 1B，α 1C， alpha1D）。 尽管其中一种克隆亚型（alpha 1D） 最初被认为是编码由基因组定义的α 1A 但最近的数据表明，这是不可能的。 的关系 必须了解克隆受体和天然亚型之间的关系， 以及在生物化学和 可以清楚地理解α 1-AR亚型的功能作用。 人SKNMC神经上皮瘤细胞表达至少两种α 1-AR， 类似于α 1A和α 1B亚型。 我们隔离了， 表达，并部分表征全长嵌合 来自与大鼠同源的该细胞系的基因组/cDNA克隆 α 1D。 我们将使用正义表达和反义敲除， SKNMC细胞中的其他人克隆，以阐明 已知克隆的药理学α 1A亚型。 如果他们是 与预期无关，我们将通过以下方法获得全长α 1A cDNA： 表达克隆 有了这些克隆人我们就能清楚地定义 药物特异性、信号传导机制和 α 1-AR亚型。 我们的主要目标包括： 1. 为了明确3个已知克隆与 α 1A亚型; 2. 通过表达获得α 1A亚型的全长cDNA， 克隆; 3. 为了确定不同的α 1-AR受体亚型 优先与不同的第二信使系统偶联;和 4. 确定不同亚型是否共存于同一细胞中 具有附加、协同或冗余的相互作用， 关于α 1-AR亚型的数量和性质的混乱。 这些实验将澄清数量，信号机制， α 1-AR亚型的相互作用。 由于药物对这些起作用 受体具有相当大的治疗重要性，这些信息可以 有直接的临床影响。