PSYCHOPHARMACOLOGICALLY SIGNIFICANT PEPTIDES

本项目的总体目标是研究 β-内啡肽（β-EP）和相关的精神药理学意义 分子水平上的肽。 具体目标如下： 研究阿片样物质的分离和表征， 来自膜制剂的非阿片样β-EP结合蛋白（受体） 在大鼠脑中，小鼠神经母细胞瘤X神经胶质瘤杂交细胞（NG 108 -15和NG 108 - 15）的细胞毒性为100%。 N18 TG 2）、人胶质母细胞瘤SF 126细胞和人神经母细胞瘤IMR-32 使用光亲和试剂2-硝基-4-叠氮基苯硫酰氯的细胞 特异性标记色氨酸残基;（B）合成两种类似物 （[Trp-4]-beta-h-EP; [D-Trp-4]-beta-h-EP），用于 β-h-EP分子中阿片样物质结构域的光标记探针。 合成[Trp-27]-beta-h-EP将用于以下物质的光标记探针： （c）为了进一步生物学表征非阿片样物质结合位点; 天然β-EP抑制肽[β-EIP，β-EP-（1-27）] 和具有抑制β-EP镇痛特性的合成类似物， 使用大鼠脑膜制备物的放射受体结合测定， 氚化二氢吗啡，[D-Ala-2，D-Leu-5]-脑啡肽， 乙基酮环唑辛、β-EP和二丙诺啡作为主要配体 通过使用豚鼠回肠、大鼠、小鼠和兔输精管进行的生物测定， （d）测试β-h-EP-（1-27）在以下中的抑制活性： 其他生物系统，如β-EP的催乳素释放活性; 从大象中分离、合成和表征β-EP 脑下垂体

A general goal of this proposed project is to investigate the function of beta-endorphin (Beta-EP) and related psychopharmacologically significant peptides at the molecular level. The specific aims are as follows: (a) To investigate on the isolation and characterization of the opioid and non-opioid beta-EP binding proteins (receptors) from membrane preparations of rat brains, mouse neuroblastoma x glioma hybrid cells (NG108-15 and N18TG2), human glioblastoma SF126 cells and human neuroblastoma IMR-32 cells using a photoaffinity reagent 2-nitro-4-azidophenylsulfenyl chloride specific to label tryptophan residue; (b) To synthesize two analogs ([Trp-4]-beta-h-EP; [D-Trp-4]-beta-h-EP) by solid-phase method to use for photolabel probes of the opioid domains in the beta-h-EP molecule. Synthetic [Trp-27]-beta-h-EP will be used for photolabel probes of non-opioid binding sites; (c) To further characterize biologically the naturally occurring beta-EP-inhibiting peptide [beta-EIP, beta-EP-(1-27)] and the synthetic analogs with inhibiting beta-EP analgesia property by radioreceptor binding assay using rat brain membrane preparations and tritiated dihydromorphine, [D-Ala-2, D-Leu-5]-enkephalin, ethylketocyclazocine, beta-EP and diprenorphine as primary ligands as well as by bioassays using guinea pig ileum, rat, mouse and rabbit vas deferens preparations; (d) to test inhibiting activities of beta-h-EP-(1-27) in other biological systems such as prolactin-releasing activity of beta-EP; and (e) to isolate, synthesize and characterize beta-EP from elephant pituitary glands.