Analytical Biochemistry 2013-04-01

A malachite green-based assay to assess glucan phosphatase activity.

Amanda R Sherwood, Bradley C Paasch, Carolyn A Worby, Matthew S Gentry

Index: Anal. Biochem. 435(1) , 54-6, (2013)

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Abstract

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.Copyright © 2012 Elsevier Inc. All rights reserved.