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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/27107

Title: A high-throughput colorimetric assay to characterize the enzyme kinetic and cellular activity of spermidine/spermine N1-acetyltransferase 1
Authors: Han-Jia Lin;Yi-Chin Lien;Chun-Hua Hsu
Contributors: NTOU:Institute of Bioscience and Biotechnology
Keywords: Activity assay;Ellman’s reagent;Enzyme kinetics;Polyamines;Spermidine/spermine N1-acetyltransferase 1
Date: 2010-12-15
Issue Date: 2011-10-21T02:22:45Z
Publisher: Analytical Biochemistry
Abstract: 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.
Relation: 407(2), pp.226–232
URI: http://ntour.ntou.edu.tw/handle/987654321/27107
Appears in Collections:[生命科學暨生物科技學系] 期刊論文

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