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A high-throughput colorimetric assay to characterize the enzyme kinetic and cellular activity of spermidine/spermine N-acetyltransferase 1 |
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| Authors: | Han-Jia Lin Yi-Chin Lien |
| Affiliation: | a Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung 20224, Taiwan b Center for Marine Bioenvironment and Biotechnology, National Taiwan Ocean University, Keelung 20224, Taiwan c Department of Agricultural Chemistry, National Taiwan University, Taipei 10617, Taiwan |
| Abstract: | Spermidine/spermine N1-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 Km 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. |
| Keywords: | Activity assay Ellman&rsquo s reagent Enzyme kinetics Polyamines Spermidine/spermine N1-acetyltransferase 1 |
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