MAPPING THE PEPTIDE BINDING SITES OF SP AND SK RECEPTORS

SP 和 SK 受体的肽结合位点图谱

• 批准号: 3418265
• 负责人: NORMAN D BOYD
• 金额: $ 19.71万
• 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 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3418265
• 关键词: SDS polyacrylamide gel electrophoresis; affinity labeling; antireceptor antibody; covalent bond; high performance liquid chromatography; inhibitor /antagonist; mass spectrometry; molecular site; neuropeptide receptor; neuropharmacology; phenylalanine analog; protein sequence; protein structure function; receptor binding; stimulant /agonist; substance K; substance P

DESCRIPTION (adapted from the applicant's abstract): The three- dimensional structure of the binding pockets of the SP and SK receptors will be defined with respect to their transmembrane alpha-helices and extracellular domains by determining the sites of covalent attachment of a series of SP and SK derivatives containing the photoreactive amino acid p-benzoylphenylalanine spaced at different positions along the amino acid sequences of the two peptides. For each photoaffinity ligand, the amino acid residue of the binding site that becomes covalently labelled will be determined by microsequencing of purified receptor fragments initially located in the primary sequence with the aid of site-specific antibodies and mass spectrometry. A comparative analysis of the similar and distinctive features of these binding domains should provide insight into the molecular basis of peptide selectivity and receptor activation. A related project will be to determine the site of attachment of a tritiated photoreactive azide derivative of the nonpeptide SP antagonist CP-96,345. An understanding of the relationship between the sites for peptide agonists and the nonpeptide antagonist may provide insight into the mechanism of the antagonist effects of this compound, and the information to be gained from the project as a whole may advance the rational design of therapeutic agents for clinical entities in which these peptides participate.

描述（根据申请人的摘要改编）：三 SP和SK受体的结合口袋的尺寸结构 将根据其跨膜α-螺旋和 通过确定共价附着的位点，细胞外域 一系列含有光电反应性氨基酸的SP和SK衍生物 沿氨基酸在不同位置间隔的P-苯甲酰苯胺 两个肽的序列。对于每个光性配体，氨基 结合位点的酸残基被共价标记为 通过纯化受体片段的微链测序确定 借助于位点特异性抗体，位于主要序列 和质谱。对类似和的比较分析 这些结合域的独特特征应提供有关 肽选择性和受体激活的分子基础。一个 相关项目将是确定附着的位点 非肽SP拮抗剂的Tritived光电反应性叠衍生物 CP-96,345。对网站之间关系的理解 肽激动剂和非肽拮抗剂可能会洞悉 该化合物的拮抗作用的机制， 从整个项目中获得的信息可能会推进 用于临床实体的治疗剂的合理设计 这些肽参与。