標題: Fluorometric assay for alcohol sulfotransferase
作者: Chen, WT
Liu, MC
Yang, YS
生物科技學系
Department of Biological Science and Technology
關鍵字: phenol sulfotransferase;alcohol sulfotransferase;PAPS;PAP
公開日期: 1-Apr-2005
摘要: A sensitive fluorometric assay was developed for alcohol sulfotransferase (AST). This was the first continuous fluorometric assay reported for AST. It used 3 '-phosphoadenosine 5 '-phosphosulfate regenerated from 3-phosphoadenosine 5 '-phosphate by a recombinant phenol sulfotransferase (PST) using 4-methylumbelliferyl sulfate as the sulfuryl group donor. The recombinant PST did not use the alcohol substrate under the designed condition, and the sensitivity for AST activity was found to be comparable to that of radioactive assay as reported in the literature. The change of fluorescence intensity of 4-methylumbelliferone corresponded directly to the amount of active AST and was sensitive enough to measure nanogram or picomole amounts of the enzyme activity. This fluorometric assay was used to determine the activities of AST as purified form and in crude extracts of pig liver, rat liver, and Escherichia coli. Some properties of human dehydroepiandrosterone sulfotransferase were determined by this method and were found to be comparable to published data. Under similar assay conditions, the contaminated activities of arylsulfatase in crude extracts were also determined. This method not only is useful for the routine and detailed kinetic study of this important class of enzymes but also has the potential for the development of a high-throughput procedure using microplate reader. (c) 2004 Elsevier Inc. All rights reserved.
URI: http://dx.doi.org/10.1016/j.ab.2004.12.016
http://hdl.handle.net/11536/13857
ISSN: 0003-2697
DOI: 10.1016/j.ab.2004.12.016
期刊: ANALYTICAL BIOCHEMISTRY
Volume: 339
Issue: 1
起始頁: 54
結束頁: 60
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