BIOCHEMICAL CHARACTERIZATION OF OPIOID BINDING SITES

描述：这个项目的主要焦点是多个阿片受体 结合部位。首席调查员长期以来一直对 识别阿片受体的多个亚型，使用各种 技术包括受体结合和止痛研究。然而，当 MU阿片受体的克隆，很明显只有一种受体是 由Mu受体基因编码，潜在的多种受体亚型 必须来自不同的机制。在上一个资助期内， 首席研究员探索了多个u受体的概念 产生于Mu受体(MOR-1)基因的选择性剪接变体。这些 概念起源于阿片类止痛的反义研究，其中 代表MOR-1不同外显子的不同反义寡核苷酸 基因对镇痛有不同的影响。这些研究是通过以下方式进一步推进的 编码多个内含子和外显子的MOR-1基因的测序。虽然 它们的生理意义尚不清楚，至少编码了 目前已分离出该受体的12个变异体。拟议的研究将 进一步探索这些潜在的受体亚型。第一个具体目标是 提供Mu受体剪接变异体的详细特征，包括 药理和解剖学研究以及新克隆的鉴定。这个 第二个目标将在ORL-1受体上进行类似的实验，识别 该受体至少有一种新的剪接变体。第三个目标将探索 MOR-1/ORL-1杂二聚体可能存在的初步研究 发现这两种受体在细胞系中的共同表达产生 单独与任一位点不同的结合部位。第四个目标将是 几种多肽配体与OPRL-1结合特性的比较 受体。

DESCRIPTION: The principal focus of this project is multiple opioid receptor binding sites. The principal investigator has a long-established interest in identifying multiple subtypes of opioid receptors, using a variety of techniques including receptor binding and analgesia studies. However, when the mu opioid receptor was cloned, it was clear that only one type of receptor was coded by the mu receptor gene, and that potential multiple receptor subtypes must arise from different mechanisms. During the previous funding period, the principal investigator has explored the concept that multiple mu receptors arise from alternative splice variants of the mu receptor (MOR-1) gene. These concepts arose originally from antisense studies of opioid analgesia, in which different antisense oligonucleotides representing different exons of the MOR-1 gene produced different effects on analgesia. The studies were furthered by sequencing of the MOR-1 gene encoding a number of introns and exons. Although their physiological significance is not yet clear, the mRNA coding for at least 12 variants of this receptor have been isolated. The proposed studies will further explore these potential receptor subtypes. The first specific aim will provide a detailed characterization of mu receptor splice variants, including pharmacological and anatomical studies and identification of new clones. The second aim will perform similar experiments on the ORL-1 receptor, identifying at least one novel splice variant of this receptor. The third aim will explore the possible existence of MOR-1/ORL-1 heterodimers, based upon the preliminary finding that co-expression of both of these receptors into cell lines produces binding sites which are different from either site alone. The fourth aim will compare the binding properties of several peptide ligands for the OPRL-1 receptor.