Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung 20224, Taiwan.
Anal Biochem. 2010 Dec 15;407(2):226-32. doi: 10.1016/j.ab.2010.07.036. Epub 2010 Aug 6.
Spermidine/spermine N(1)-acetyltransferase 1 (SSAT1) is a key enzyme that catalyzes the catabolism of polyamines. SSAT1 is a very important enzyme because it not only maintains the homeostasis of polyamines but also is involved in many physiological and pathological events. As such, a rapid assay of SSAT1 activity is valuable in drug screening and clinical diagnostics. Here, we report a novel colorimetric assay for monitoring SSAT1 activity in zebrafish (zSSAT1). In comparison with the available SSAT1 assays, this new method is cost-effective and simple. The optimal zSSAT1 activity was obtained below 55°C in a mild alkaline environment. The K(m) values of zSSAT1 for spermidine and spermine are 55 and 182 μM, respectively, whereas putrescine is not a good substrate for zSSAT1. In addition to enzyme kinetic studies, the colorimetric assay was also used to detect the cellular activity of SSAT1. Thus, the current method is a reliable assay for determining SSAT1 activity with many potential applications in medical biology.
精脒/精胺 N(1)-乙酰基转移酶 1（SSAT1）是一种关键的酶，可催化多胺的分解代谢。SSAT1 是一种非常重要的酶，因为它不仅维持多胺的内稳态，而且还参与许多生理和病理事件。因此，快速测定 SSAT1 活性在药物筛选和临床诊断中具有重要价值。在这里，我们报道了一种用于监测斑马鱼（zSSAT1）中 SSAT1 活性的新型比色测定法。与现有的 SSAT1 测定法相比，这种新方法具有成本效益和简单的特点。在温和的碱性环境中，最佳的 zSSAT1 活性在 55°C 以下获得。zSSAT1 对亚精胺和精胺的 K(m) 值分别为 55 和 182μM，而腐胺不是 zSSAT1 的良好底物。除了酶动力学研究外，比色测定法还用于检测 SSAT1 的细胞活性。因此，目前的方法是一种可靠的测定 SSAT1 活性的方法，在医学生物学中有许多潜在的应用。
