Department of Molecular and Cellular Biochemistry, Center for Structural Biology, University of Kentucky, Lexington, KY 40536, USA.
Anal Biochem. 2013 Apr 1;435(1):54-6. doi: 10.1016/j.ab.2012.10.044. Epub 2012 Nov 29.
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.
最近发现了一类独特的双特异性磷酸酶，它们可以使葡聚糖去磷酸化，因此我们报告了一种针对磷酸化葡聚糖的磷酸酶活性检测的体外测定法。我们证明，与使用合成底物的一般磷酸酶测定法相反，只有具有葡聚糖磷酸酶活性的磷酸酶才能使用所述测定法从磷酸化的支链淀粉中释放出磷酸盐。该测定法简单且具有成本效益，可提供可重复的结果，明确确定是否存在葡聚糖磷酸酶活性。所描述的测定法将成为表征葡聚糖磷酸酶家族中新出现成员的有用工具。
