Joh T H, Shikimi T, Pickel V M, Reis D J
Proc Natl Acad Sci U S A. 1975 Sep;72(9):3575-9. doi: 10.1073/pnas.72.9.3575.
Tryptophan hydroxylase [EC 1.14.16.4; L-tryptophan, tetrahydropteridine:oxygen oxidoreductase (5-hydroxylating)], the enzyme catalyzing the rate-limiting step in the biosynthesis of serotonin, was purified 79-fold from the region of the raphe nucleus of rat midbrain by sequential column chromatography and disc-gel electrophoresis. In electrophoresis three bands were distinguished, A, B, and C, which, when separated and submitted individually to electrophoresis, reproduced the same three bands. Bands A and C were enzymatically active and inhibited by para-chlorohenylalanine. Antibodies produced to each of the three bands crossreacted by immuno double diffusion and electrophoresis with each other and homogenates of raphe nuclei; they completely inhibited enzyme activity only of tryptophan hydroxylase. Tryptophan hydroxylase was localized by light and electron immunohistochemistry to serotonin neutrons of the raphe. Ultrastructurally, in cell bodies, the enzyme was distributed in cytoplasm and in association with endoplasmic reticulum and Golgi apparatus. In dendrites and axons, it was associated with microtubules. Tryptophan hydroxylase in brain is only neuronal and cytoplasmic, exists in multiple forms, and is associated with microtubules, suggesting it may be transported from sites of synthesis in cell body into axons.
色氨酸羟化酶[EC 1.14.16.4；L-色氨酸，四氢蝶啶：氧氧化还原酶（5-羟化）]，催化血清素生物合成限速步骤的酶，通过连续柱色谱法和圆盘凝胶电泳从大鼠中脑缝际核区域纯化了79倍。在电泳中可区分出三条带，A、B和C，当将它们分离并分别进行电泳时，会重现相同的三条带。带A和C具有酶活性，并被对氯苯丙氨酸抑制。针对这三条带各自产生的抗体通过免疫双扩散和电泳相互交叉反应，并与缝际核匀浆交叉反应；它们仅完全抑制色氨酸羟化酶的酶活性。通过光镜和电镜免疫组织化学将色氨酸羟化酶定位到缝际的血清素神经元。在超微结构上，在细胞体中，该酶分布于细胞质中，并与内质网和高尔基体相关。在树突和轴突中，它与微管相关。脑中的色氨酸羟化酶仅存在于神经元和细胞质中，以多种形式存在，并与微管相关，表明它可能从细胞体中的合成部位转运到轴突中。
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