A malachite green-based assay to assess glucan phosphatase activity
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...
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| Veröffentlicht in: | Analytical biochemistry 2013-04, Vol.435 (1), p.54-56 |
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| container_title | Analytical biochemistry |
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| creator | Sherwood, Amanda R. Paasch, Bradley C. Worby, Carolyn A. Gentry, Matthew S. |
| description | 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. |
| doi_str_mv | 10.1016/j.ab.2012.10.044 |
| format | Article |
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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.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/j.ab.2012.10.044</identifier><identifier>PMID: 23201267</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amylopectin ; Amylopectin - metabolism ; Animals ; Arabidopsis - enzymology ; cost effectiveness ; Dual-Specificity Phosphatases - metabolism ; Enzyme Assays - economics ; Enzyme Assays - methods ; Glucan ; Glycogen ; Glycogen - metabolism ; Humans ; in vitro studies ; malachite ; Malachite green ; Phosphatase ; phosphates ; Phosphorylation ; Rabbits ; Rosaniline Dyes - analysis ; Rosaniline Dyes - metabolism ; Starch ; Starch - metabolism ; Substrate Specificity</subject><ispartof>Analytical biochemistry, 2013-04, Vol.435 (1), p.54-56</ispartof><rights>2012 Elsevier Inc.</rights><rights>Copyright © 2012 Elsevier Inc. All rights reserved.</rights><rights>2012 Elsevier Inc. All rights reserved. 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c570t-3fd702e6e0d3b7868af8807240027339b6c5aca5aeb056d3dd64c7cf473782753</citedby><cites>FETCH-LOGICAL-c570t-3fd702e6e0d3b7868af8807240027339b6c5aca5aeb056d3dd64c7cf473782753</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0003269712005787$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,776,780,881,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23201267$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sherwood, Amanda R.</creatorcontrib><creatorcontrib>Paasch, Bradley C.</creatorcontrib><creatorcontrib>Worby, Carolyn A.</creatorcontrib><creatorcontrib>Gentry, Matthew S.</creatorcontrib><title>A malachite green-based assay to assess glucan phosphatase activity</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>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.</description><subject>Amylopectin</subject><subject>Amylopectin - metabolism</subject><subject>Animals</subject><subject>Arabidopsis - enzymology</subject><subject>cost effectiveness</subject><subject>Dual-Specificity Phosphatases - metabolism</subject><subject>Enzyme Assays - economics</subject><subject>Enzyme Assays - methods</subject><subject>Glucan</subject><subject>Glycogen</subject><subject>Glycogen - metabolism</subject><subject>Humans</subject><subject>in vitro studies</subject><subject>malachite</subject><subject>Malachite green</subject><subject>Phosphatase</subject><subject>phosphates</subject><subject>Phosphorylation</subject><subject>Rabbits</subject><subject>Rosaniline Dyes - analysis</subject><subject>Rosaniline Dyes - metabolism</subject><subject>Starch</subject><subject>Starch - metabolism</subject><subject>Substrate Specificity</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkT1v2zAQhomiReOm3Tu1GrvIOZISSXUoEBj9CBCgQ5KZOFEnm4YsuSRtwP8-FJwG7VCUC3m85z0e72XsPYclB66utktslwK4yOESquoFW3BoVAkSmpdsAQCyFKrRF-xNjFsAzqtavWYXQs4ipRdsdV3scEC38YmKdSAayxYjdQXGiKciTfOBYizWw8HhWOw3U9xvMGWmQJf80afTW_aqxyHSu6f9kj18-3q_-lHe_vx-s7q-LV2tIZWy7zQIUgSdbLVRBntjQIsKQGgpm1a5Gh3WSC3UqpNdpyqnXV9pqY3QtbxkX85194d2R52jMQUc7D74HYaTndDbvzOj39j1dLSy1rqpq1zg01OBMP06UEx256OjYcCRpkO0vJmXqYz6PyqMaZo8TZNROKMuTDEG6p874mBnm-zWYmvnic832aYs-fDnT54Fv33JwMcz0ONkcR18tA93Oadg9lSpucHPZ4LyxI-ego3O0-io84Fcst3k__3-I8SDqu4</recordid><startdate>20130401</startdate><enddate>20130401</enddate><creator>Sherwood, Amanda R.</creator><creator>Paasch, Bradley C.</creator><creator>Worby, Carolyn A.</creator><creator>Gentry, Matthew S.</creator><general>Elsevier Inc</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20130401</creationdate><title>A malachite green-based assay to assess glucan phosphatase activity</title><author>Sherwood, Amanda R. ; Paasch, Bradley C. ; Worby, Carolyn A. ; Gentry, Matthew S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c570t-3fd702e6e0d3b7868af8807240027339b6c5aca5aeb056d3dd64c7cf473782753</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Amylopectin</topic><topic>Amylopectin - metabolism</topic><topic>Animals</topic><topic>Arabidopsis - enzymology</topic><topic>cost effectiveness</topic><topic>Dual-Specificity Phosphatases - metabolism</topic><topic>Enzyme Assays - economics</topic><topic>Enzyme Assays - methods</topic><topic>Glucan</topic><topic>Glycogen</topic><topic>Glycogen - metabolism</topic><topic>Humans</topic><topic>in vitro studies</topic><topic>malachite</topic><topic>Malachite green</topic><topic>Phosphatase</topic><topic>phosphates</topic><topic>Phosphorylation</topic><topic>Rabbits</topic><topic>Rosaniline Dyes - analysis</topic><topic>Rosaniline Dyes - metabolism</topic><topic>Starch</topic><topic>Starch - metabolism</topic><topic>Substrate Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sherwood, Amanda R.</creatorcontrib><creatorcontrib>Paasch, Bradley C.</creatorcontrib><creatorcontrib>Worby, Carolyn A.</creatorcontrib><creatorcontrib>Gentry, Matthew S.</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sherwood, Amanda R.</au><au>Paasch, Bradley C.</au><au>Worby, Carolyn A.</au><au>Gentry, Matthew S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A malachite green-based assay to assess glucan phosphatase activity</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>2013-04-01</date><risdate>2013</risdate><volume>435</volume><issue>1</issue><spage>54</spage><epage>56</epage><pages>54-56</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><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.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>23201267</pmid><doi>10.1016/j.ab.2012.10.044</doi><tpages>3</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Analytical biochemistry, 2013-04, Vol.435 (1), p.54-56 |
| issn | 0003-2697 1096-0309 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Amylopectin Amylopectin - metabolism Animals Arabidopsis - enzymology cost effectiveness Dual-Specificity Phosphatases - metabolism Enzyme Assays - economics Enzyme Assays - methods Glucan Glycogen Glycogen - metabolism Humans in vitro studies malachite Malachite green Phosphatase phosphates Phosphorylation Rabbits Rosaniline Dyes - analysis Rosaniline Dyes - metabolism Starch Starch - metabolism Substrate Specificity |
| title | A malachite green-based assay to assess glucan phosphatase activity |
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