Bohmaker K, Puza T, Goldstein M, Meller E
Millhauser Laboratories, Department of Psychiatry, University School of Medicine, New York, New York.
J Pharmacol Exp Ther. 1989 Jan;248(1):97-103.
Incubation of rat striatal slices with forskolin (0.05-10 microM) elicited a dose-dependent increase in the activity of tyrosine hydroxylase (TH) assayed in subsequently solubilized extracts of the enzyme. At low concentrations (33 microM) of the cofactor (6R)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride TH activity was increased 2.5 to 3-fold. Kinetic analysis of TH activity as a function of (6R)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride concentration indicated that the enzyme isolated from control slices was composed of multiple species with different Km's for cofactor. Treatment with forskolin (1.5-15 microM) converted the enzyme into a single species with a low Km (28 microM) for (6R)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride. The dopamine (DA) agonist R-(-)-N-n-propylnorapomorphine (0.1 microM) reversed forskolin-induced activation of TH. Concentration-response curves were obtained for inhibition of forskolin-stimulated TH by R-(-)-N-n-propylnorapomorphine and the DA autoreceptor-selective agonists (+)- and (-)-3-(3-hydroxyphenyl)-N-n-propylpiperidine and 3-[4-(4-phenyl-1,2,3,6-tetrahydropyridyl-1)-butyl]indole. R-(-)-N-n-propylnorapomorphine maximally inhibited forskolin-stimulated activity 85%, as indicated by ALLFIT computer analysis of concentration-response curves. (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine and 3-[4-(4-phenyl-1,2,3,6-tetrahydropyridyl-1)-butyl]indole produced a lower degree of maximal inhibition (54 and 63%, respectively), whereas (-)-3-(3-hydroxyphenyl)-N-n-propylpiperidine was inactive. The D2 DA receptor blocker sulpiride (1 microM) competitively antagonised the effects of all the agonists.(ABSTRACT TRUNCATED AT 250 WORDS)
用福司可林(0.05 - 10微摩尔)孵育大鼠纹状体切片,可使随后对该酶进行溶解提取后所检测的酪氨酸羟化酶(TH)活性呈剂量依赖性增加。在低浓度(33微摩尔)的辅因子(6R)-5,6,7,8-四氢-L-生物蝶呤二盐酸盐存在下,TH活性增加2.5至3倍。对TH活性作为(6R)-5,6,7,8-四氢-L-生物蝶呤二盐酸盐浓度的函数进行动力学分析表明,从对照切片中分离出的该酶由多种对辅因子具有不同米氏常数(Km)的物种组成。用福司可林(1.5 - 15微摩尔)处理后,该酶转变为对(6R)-5,6,7,8-四氢-L-生物蝶呤二盐酸盐具有低Km(28微摩尔)的单一物种。多巴胺(DA)激动剂R-(-)-N-正丙基去甲阿朴吗啡(0.1微摩尔)可逆转福司可林诱导的TH激活。获得了R-(-)-N-正丙基去甲阿朴吗啡以及DA自受体选择性激动剂(+)-和(-)-3-(3-羟基苯基)-N-正丙基哌啶和3-[4-(4-苯基-1,2,3,6-四氢吡啶基-1)-丁基]吲哚对福司可林刺激的TH抑制作用的浓度-反应曲线。如对浓度-反应曲线进行ALLFIT计算机分析所示,R-(-)-N-正丙基去甲阿朴吗啡最大程度地抑制福司可林刺激的活性达85%。(+)-3-(3-羟基苯基)-N-正丙基哌啶和3-[4-(4-苯基-1,2,3,6-四氢吡啶基-1)-丁基]吲哚产生的最大抑制程度较低(分别为54%和63%),而(-)-3-(3-羟基苯基)-N-正丙基哌啶无活性。D2 DA受体阻滞剂舒必利(1微摩尔)竞争性拮抗所有激动剂的作用。(摘要截短至250字)