PSYCHOPHARMACOLOGICALLY SIGNIFICANT PEPTIDES

这个提议的项目的一个总体目标是研究 β-内啡肽(β-EP)及其相关的精神药理学意义 分子水平上的多肽。具体目标如下：(A) 阿片类药物的分离与鉴定研究 膜制剂中的非阿片类β-EP结合蛋白(受体) 大鼠脑、小鼠神经母细胞瘤×脑胶质瘤杂交细胞(NG108-15和 N18TG2)、人胶质母细胞瘤SF126细胞和人神经母细胞瘤IMR-32 使用光亲和试剂2-硝基-4-叠氮苯硫酰氯的细胞 特异性标记色氨酸残基；(B)合成两个类似物 ([Trp-4]-β-h-EP；[D-Trp-4]-beta-h-EP) β-h-EP分子中阿片结构域的光标记探针。 合成的[Trp-27]-β-h-EP将用于 非阿片类药物结合部位；(C)进一步确定 天然β-EP抑制肽[β-EIP，β-EP-(1-27)] 以及合成的抑制β-EP镇痛作用的类似物 大鼠脑膜制剂与放射受体结合分析 氚二氢吗啡，[D-丙氨酸-2，D-亮氨酸-5]-脑啡肽， 乙酮环氮唑、β-EP和异丙诺啡也是主要配体 用豚鼠回肠、大鼠、小鼠和兔输精管进行生物测定 制剂；(D)测试β-h-EP-(1-27)在 其他生物系统，如催乳素释放活性的β-EP； 和(E)从大象中分离、合成和鉴定β-EP 脑垂体腺。

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.