A malachite green-based assay to assess glucan phosphatase activity

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• 作者：Amanda R. Sherwood ; Bradley C. Paasch ; Carolyn A. Worby ; Matthew S. Gentry ; ^{matthew.gentry@uky.edu}
• 关键词：Glucan ; Phosphatase ; Malachite green ; Amylopectin ; Glycogen ; Starch
• 刊名：Analytical Biochemistry
• 出版年：2013
• 出版时间：1 April, 2013
• 年：2013
• 卷：435
• 期：1
• 页码：54-56
• 全文大小：289 K

文摘

With the recent discovery of a unique class of dual-specificity phosphatases that dephosphorylate glucans, we report an in vitro assay tailored for the detection of phosphatase activity against phosphorylated glucans. We demonstrate that, in contrast to a general phosphatase assay using a synthetic substrate, only phosphatases that possess glucan phosphatase activity liberate phosphate from the phosphorylated glucan amylopectin using the described assay. This assay is simple and cost-effective, providing reproducible results that clearly establish the presence or absence of glucan phosphatase activity. The assay described will be a useful tool in characterizing emerging members of the glucan phosphatase family.