title: A malachite green-based assay to assess glucan phosphatase activity.
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.
authors.affiliation: Department of Molecular and Cellular Biochemistry, Center for Structural Biology, University of Kentucky, Lexington, KY 40536, USA.
authors.forename: Amanda R
authors.initials: AR
authors.lastname: Sherwood
authors.forename: Bradley C
authors.initials: BC
authors.lastname: Paasch
authors.forename: Carolyn A
authors.initials: CA
authors.lastname: Worby
authors.forename: Matthew S
authors.initials: MS
authors.lastname: Gentry
mesh_terms: D000687:Amylopectin / Q000378:metabolism*; D000818:Animals; D017360:Arabidopsis / Q000201:enzymology; D054637:Dual-Specificity Phosphatases / Q000378:metabolism*; D057075:Enzyme Assays / Q000191:economics / Q000379:methods*; D006003:Glycogen / Q000378:metabolism; D006801:Humans; D010766:Phosphorylation; D011817:Rabbits; D012394:Rosaniline Dyes / Q000032:analysis* / Q000378:metabolism; D013213:Starch / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D012394:Rosaniline Dyes; C005095:malachite green; D013213:Starch; D006003:Glycogen; D000687:Amylopectin; D054637:Dual-Specificity Phosphatases