SP RECEPTOR BINDING SITES FOR AGONISTS AND ANTAGONISTS

激动剂和拮抗剂的 SP 受体结合位点

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/2839354
• 关键词: CHO cells; SDS polyacrylamide gel electrophoresis; affinity labeling; biological signal transduction; crosslink; disulfide bond; high performance liquid chromatography; immunoaffinity chromatography; immunoprecipitation; mass spectrometry; matrix assisted laser desorption ionization; molecular site; neuropeptide receptor; neurotransmitter agonist; neurotransmitter antagonist; posttranslational modifications; protein sequence; protein structure function; receptor binding; receptor coupling; site directed mutagenesis; substance P; transcription factor; western blottings

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研究的延续

项目成果

Molecular characterization of the substance P*neurokinin-1 receptor complex: development of an experimentally based model.

P*neurokinin-1 受体复合物物质的分子表征：基于实验的模型的开发。

• DOI: 10.1074/jbc.m101057200
• 发表时间: 2001
• 期刊: The Journal of biological chemistry
• 作者: Pellegrini,M;Bremer,AA;Ulfers,AL;Boyd,ND;Mierke,DF
• 通讯作者: Mierke,DF

Evidence for spatial proximity of two distinct receptor regions in the substance P (SP)*neurokinin-1 receptor (NK-1R) complex obtained by photolabeling the NK-1R with p-benzoylphenylalanine3-SP.

P 物质 (SP)*神经激肽-1 受体 (NK-1R) 复合物中两个不同受体区域空间接近的证据，通过用对苯甲酰基苯丙氨酸 3-SP 光标记 NK-1R 获得。

• DOI: 10.1074/jbc.m100824200
• 发表时间: 2001
• 期刊: The Journal of biological chemistry
• 作者: Bremer,AA;Leeman,SE;Boyd,ND
• 通讯作者: Boyd,ND

Identification of the site in the substance P (NK-1) receptor for modulation of peptide binding by sulfhydryl reagents.

识别 P 物质 (NK-1) 受体中用于通过巯基试剂调节肽结合的位点。

• DOI: 10.1074/jbc.271.4.1950
• 发表时间: 1996
• 期刊: The Journal of biological chemistry
• 作者: Li,H;Hsu,P;Sachais,BS;Krause,JE;Leeman,SE;Boyd,ND
• 通讯作者: Boyd,ND

Direct evidence for the interaction of neurokinin A with the tachykinin NK(1) receptor in tissue.

神经激肽 A 与组织中速激肽 NK(1) 受体相互作用的直接证据。

• DOI: 10.1016/s0014-2999(01)01107-4
• 发表时间: 2001
• 期刊: European journal of pharmacology
• 影响因子: 5
• 作者: Bremer,AA;Tansky,MF;Wu,M;Boyd,ND;Leeman,SE
• 通讯作者: Leeman,SE

Photoaffinity labeling of mutant neurokinin-1 receptors reveals additional structural features of the substance P/NK-1 receptor complex.

突变型神经激肽-1 受体的光亲和标记揭示了 P/NK-1 受体复合物物质的其他结构特征。

• DOI: 10.1021/bi001880x
• 发表时间: 2001
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Macdonald,D;Mierke,DF;Li,H;Pellegrini,M;Sachais,B;Krause,JE;Leeman,SE;Boyd,ND
• 通讯作者: Boyd,ND