Reactive affinity probes for the mapping of the glycine-binding site of the NMDA receptor NR1 subunit

作     者：Kreimeyer, A Laube, B Sturgess, M Goeldner, M Foucaud, B 

作者机构：ULP Fac Pharm Chim Bioorgan Lab CNRS UMR 7514 F-67401 Illkirch Graffenstaden France Max Planck Inst Brain Res D-60528 Frankfurt Germany Bearsden Bio Inc Aston PA 19014 USA 

出 版 物：《JOURNAL OF RECEPTOR AND SIGNAL TRANSDUCTION RESEARCH》 (受体与信号转导杂志)

年 卷 期：1999年第19卷第1-4期

页      面：547-557页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 09[农学] 

主　　题：亲和力标记物/化学合成 亲和力标记物/化学 亲和力标记物/代谢 结合部位 脑/代谢 兴奋性氨基酸拮抗剂/化学合成 兴奋性氨基酸拮抗剂/化学 兴奋性氨基酸拮抗剂/代谢 甘氨酸/化学 动力学 配体 分子结构 蛋白质构象 喹诺酮类/化学 喹诺酮类/代谢 喹诺酮类/药理学 动物 大鼠 

摘      要：The glycine co-agonist binding site of the NMDA receptor is a target for the prevention and treatment of neurotoxic and neurodegenerative conditions. Until now, the interactions taking place at this site, and its structure, have been investigated by ligand structure-activity relationships and by site-directed mutagenesis. On the basis of a structural model which is currently proposed for this site, we have designed and synthesized six affinity markers by substituting electrophilic reactive groups in the 4, the 7 and the 3 positions of L 701,324, a high-affinity glycine site antagonist. These compounds compete with H-3-DCKA binding to rat brain membranes at equilibrium with nanomolar to low-micromolar affinities, and antagonize glycine-evoked currents in oocytes transfected with wild-type NR1-NR2B. However, they do not induce a time-shift in binding equilibria, and do not inactivate irreversibly the glycine evoked currents. Since they react only with cysteine at physiological pH, we conclude that there is no such residue in the site, in agreement with the model. Our affinity markers therefore represent potential topological probes for NMDA receptors with sequence positions related to the glycine-binding site mutated into cysteine.