SP RECEPTOR BINDING SITES FOR AGONISTS AND ANTAGONISTS

激动剂和拮抗剂的 SP 受体结合位点

基本信息

批准号：
2839354
负责人：
NORMAN D BOYD
金额：
$ 28.13万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-12-01 至 2001-06-30
项目状态：
已结题

项目摘要

This renewal is a continuation of studies on the structure and function of the substance P (SP) receptor. The long term objective of this proposal is to understand, at the molecular level, the basis of agonist and antagonist specificity, receptor activation and downstream signaling events. Our approach will be to use photoreactive SP analogs containing p-benzoylphenylalanine substituted at different positions in the eleven amino acid sequence of SP. The site(s) of covalent attachment of these photoligands on the SP receptor will be determined by MALDI-mass spectrometric analysis of isolated photolabeled receptor fragments, which are generated by enzymatic and/or chemical cleavage of the receptor. These studies will aid in defining the SP binding pocket and orient the peptide within that pocket. We will develop benzophenone-containing derivatives of non-peptide SP antagonists as photoprobes of the antagonist binding domain(s). An understanding of the relationship between the binding sites for peptide agonist and the non-peptide antagonists forms the foundation for understanding underlying mechanisms of antagonism, which is essential for drug development. The availability of photoreactive peptide agonists, together with chemical crosslinking and immunodetection, provides us with tools to identify specific G proteins and other regulatory proteins which interact with the SP receptor. This approach will provide important information about the signal transduction machinery associated with the SP receptor. As it has been estimated that up to 60 percent of all medicines used today exert their effects through G protein signaling pathways, these studies may provide new and interesting information of basic clinical relevance. The peptide substance P has attracted considerable attention because of its multiple biological activities: as a neurotransmitter in the central, sensory and autonomic nervous systems; as an agent that causes contraction of smooth muscle in the gastrointestinal tract and as a mediator of inflammation and immune responses. SP has been implicated in a number of sensory and neurogenerative disorders. The information provided by our continuing effort should form the basis for understanding and development of novel SP receptor agonists and antagonists.

这种更新是对结构和功能的研究的延续物质p（sp）受体。这个长期目标建议是在分子水平上理解激动剂的基础以及拮抗剂特异性，受体激活和下游信号传导事件。我们的方法是使用光电反应性SP类似物含有在不同位置取代的p-苯甲酰丙氨酸 Sp的11个氨基酸序列。共价站点将确定这些光团在SP受体上的附着通过MALDI-MAS光谱分析分离的光标记受体片段，由酶和/或化学产生受体的切割。这些研究将有助于定义SP 绑定口袋并将肽定向在该口袋内。我们将发展非肽SP的含苯甲酮的衍生物拮抗剂作为拮抗剂结合结构域的光探测。一个了解肽的结合位点之间的关系激动剂和非肽拮抗剂构成了了解拮抗的潜在机制，这是必不可少的用于药物开发。光电反应性肽的可用性激动剂以及化学交联和免疫试验，为我们提供了识别特定G蛋白和其他的工具与SP受体相互作用的调节蛋白。这种方法将提供有关信号转导的重要信息与SP受体相关的机械。据估计当今使用的所有药物中，多达60％的效果通过G蛋白信号通路，这些研究可能提供新的以及基本临床相关性的有趣信息。肽物质P引起了很大的关注它的多种生物学活动：作为中央的神经递质，感觉和自主神经系统；作为导致的代理胃肠道中平滑肌的收缩，作为一个炎症和免疫反应的介体。 SP一直是与许多感觉和神经发生疾病有关。这我们持续努力提供的信息应构成理解和开发新型SP受体激动剂和对手。