SYNTHESIS AND RELEASE OF BIOGENIC AMINES

这些研究集中在酪氨酸的调节特性 羟化酶、色氨酸羟化酶、一氧化氮合酶（NOS）和 它们的辅因子四氢生物蝶呤（BH 4）在生物合成和释放 脑中的儿茶酚胺和血清素 我们已经证实，一氧化氮（NO）来源于L-精氨酸，通过 NOS的作用增强纹状体切片中多巴胺的释放。 这 NOS促神经生长剂-硝基精氨酸可抑制这种作用。 我们有 也证实了BH 4也能增强纹状体多巴胺的释放， 切片 我们的研究表明，L-精氨酸的刺激作用， 增强多巴胺释放可以减少在切片中， 细胞内BH 4水平已经耗尽。 BH 4消耗和 抑制NOS同样有效地阻断L-精氨酸- 介导的多巴胺释放刺激。 这一发现支持了 结论L-精氨酸介导的多巴胺释放刺激 依赖BH 4。 我们已经从大鼠和牛脑中纯化了14-3-3蛋白， 研究了14-3-3在人重组 色氨酸羟化酶（与麦芽糖结合蛋白融合）， E.coli. 融合色氨酸羟化酶磷酸化， cAMP依赖性蛋白激酶和钙调蛋白依赖性蛋白 激酶II。 初步结果表明，使用天然的凝胶迁移试验， 聚丙烯酰胺凝胶表明没有形成络合物。 亲和力 其中融合酶共价偶联的色谱实验 与Affi凝胶-10的活化酯的结合表明14-3-3 与偶联的酶。

These studies have focused on the regulatory properties of tyrosine hydroxylase, tryptophan hydroxylase, nitric oxide synthase (NOS) and their cofactor tetrahydrobiopterin (BH4) in the biosynthesis and release of catecholamine and serotonin in the brain. We have confirmed that nitric oxide (NO) derived from L-arginine through the action of NOS enhances dopamine release in striatal slices. This effect can be inhibited by the NOS inhibitor-nitro arginine. We have also confirmed that BH4 also enhances dopamine release in striatal slices. Our studies show that the stimulating effect of L-arginine in enhancing dopamine release can be decreased in the slices in which the intracellular BH4 levels have been depleted. BH4 depletion and inhibition of NOS are equally effective in blocking the L-arginine- mediated stimulation of dopamine release. This finding supports the conclusion that the L-arginine-mediated stimulation of dopamine release is BH4 dependent. We have purified the 14-3-3 protein from rat and bovine brains and investigated the role of 14-3-3 in the regulation of human recombinant tryptophan hydroxylase (fused with maltose binding protein) expressed in E.coli. Fusion tryptophan hydroxylase was phosphorylated in the presence of both cAMP-dependent protein kinase and calmodulin-dependent protein kinase II. Preliminary results showed that gel shift assay using native polyacrylamide gel indicated no complex formation. Affinity chromatography experiments where fusion enzyme was covalently coupled with the activated ester of Affi gel-10 indicated no binding of 14-3-3 with the coupled enzyme.