Chae H D, Park T J, Lee Y K, Lee T G, Kim K T
Department of Life Science, Pohang University of Science and Technology, South Korea.
Neurochem Int. 1999 Dec;35(6):447-51. doi: 10.1016/s0197-0186(99)00086-8.
We report here a rapid, simple, and accurate method to assay for serotonin N-acetyltransferase (NAT) activity. This assay relies on the selective diffusion of radiolabeled acetyltryptamine into a water-immiscible scintillation fluid. Unlike organic solvent extraction, thin-layer chromatography, or high performance liquid chromatography, the separation of acetyltryptamine from acetyl CoA and tryptamine is not required in the method. Moreover, the limit of sensitivity is less than 4 pmol of N-acetyltryptamine formed per sample. Enhancement of NAT activity upon beta-adrenergic receptor stimulation in the rat pineal gland was clearly detected with this method. In addition, the NAT activity measurements obtained with this method agreed quantitatively in the pineal gland and other brain tissues with the conventional organic solvent extraction method. The results suggest that this liquid biphasic diffusion assay is applicable to the detection of NAT activity in tissues and cells.
我们在此报告一种快速、简便且准确的测定血清素N - 乙酰转移酶（NAT）活性的方法。该测定法依赖于放射性标记的乙酰色胺选择性扩散到与水不混溶的闪烁液中。与有机溶剂萃取、薄层色谱法或高效液相色谱法不同，该方法不需要将乙酰色胺与乙酰辅酶A和色胺分离。此外，灵敏度极限为每个样品形成的N - 乙酰色胺小于4皮摩尔。用该方法可清晰检测到大鼠松果体中β - 肾上腺素能受体刺激后NAT活性的增强。另外，用该方法获得的NAT活性测量值在松果体和其他脑组织中与传统的有机溶剂萃取法在定量上是一致的。结果表明，这种液 - 液双相扩散测定法适用于检测组织和细胞中的NAT活性。
