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Anal Biochem. 2013 Apr 1;435(1):54-6. doi: 10.1016/j.ab.2012.10.044. Epub 2012 Nov 29.

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

Author information

1
Department of Molecular and Cellular Biochemistry, Center for Structural Biology, University of Kentucky, Lexington, KY 40536, USA.

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.

PMID:
23201267
PMCID:
PMC3577954
DOI:
10.1016/j.ab.2012.10.044
[Indexed for MEDLINE]
Free PMC Article

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