Determination of the starch-phosphorylating enzyme activity in plant extracts
For quantification of α-glucan, water dikinase (GWD) activity in crude extracts of plant tissues a radio-labeling assay was established that uses soluble starch and 33P-labeled ATP as phosphate acceptor and donor, respectively. A constant rate of starch labeling was observed only if the ATP applied...
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| Veröffentlicht in: | Planta 2003-03, Vol.216 (5), p.798-801 |
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| creator | Ritte, Gerhard Steup, Martin Kossmann, Jens Lloyd, James R. |
| description | For quantification of α-glucan, water dikinase (GWD) activity in crude extracts of plant tissues a radio-labeling assay was established that uses soluble starch and 33P-labeled ATP as phosphate acceptor and donor, respectively. A constant rate of starch labeling was observed only if the ATP applied was labeled at the β position. In wild-type extracts from leaves of Arabidopsis thaliana (L.) Heynh. the maximum rate of starch phosphorylation was approximately 27 pmol min-1 (mg protein)-1. Leaf extracts from the GWD-deficient sex1 mutants of Arabidopsis showed no significant incorporation of phosphate whereas extracts from potato (Solanum tuberosum L.) tuber expressing a GWD antisense construct exhibited less activity than the wild-type control. To our knowledge this is the first time that a quantification of the starch-phosphorylating activity has been achieved in plant crude extracts. |
| doi_str_mv | 10.1007/s00425-002-0931-1 |
| format | Article |
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A constant rate of starch labeling was observed only if the ATP applied was labeled at the β position. In wild-type extracts from leaves of Arabidopsis thaliana (L.) Heynh. the maximum rate of starch phosphorylation was approximately 27 pmol min-1 (mg protein)-1. Leaf extracts from the GWD-deficient sex1 mutants of Arabidopsis showed no significant incorporation of phosphate whereas extracts from potato (Solanum tuberosum L.) tuber expressing a GWD antisense construct exhibited less activity than the wild-type control. To our knowledge this is the first time that a quantification of the starch-phosphorylating activity has been achieved in plant crude extracts.</description><identifier>ISSN: 0032-0935</identifier><identifier>EISSN: 1432-2048</identifier><identifier>DOI: 10.1007/s00425-002-0931-1</identifier><identifier>PMID: 12624767</identifier><identifier>CODEN: PLANAB</identifier><language>eng</language><publisher>Berlin: Springer-Verlag</publisher><subject>Adenosine Triphosphate - metabolism ; Alleles ; Arabidopsis - enzymology ; Arabidopsis Proteins - metabolism ; ATP ; Biological and medical sciences ; Crop harvesting ; Enzymatic activity ; Enzyme activity ; Fundamental and applied biological sciences. Psychology ; Glucans - metabolism ; Leaves ; Metabolism ; Monosaccharide Transport Proteins - metabolism ; Phosphates ; Phosphorus Radioisotopes ; Phosphorylation ; Photosynthesis, respiration. Anabolism, catabolism ; Plant extracts ; Plant Extracts - isolation & purification ; Plant Extracts - metabolism ; Plant Leaves - enzymology ; Plant physiology and development ; Plant tissues ; Plants ; Random allocation ; Solanum tuberosum - enzymology ; Starch ; Starch - metabolism ; Starch Phosphorylase - metabolism ; Starches ; Tubers</subject><ispartof>Planta, 2003-03, Vol.216 (5), p.798-801</ispartof><rights>Springer-Verlag 2002</rights><rights>2003 INIST-CNRS</rights><rights>Springer-Verlag 2003</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c376t-4aa4f5c7a372f5652a2085e7e6a4e7489b54cbdb848e93341ac47278930028253</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/23387377$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/23387377$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>315,781,785,804,27929,27930,58022,58255</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14616009$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12624767$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ritte, Gerhard</creatorcontrib><creatorcontrib>Steup, Martin</creatorcontrib><creatorcontrib>Kossmann, Jens</creatorcontrib><creatorcontrib>Lloyd, James R.</creatorcontrib><title>Determination of the starch-phosphorylating enzyme activity in plant extracts</title><title>Planta</title><addtitle>Planta</addtitle><description>For quantification of α-glucan, water dikinase (GWD) activity in crude extracts of plant tissues a radio-labeling assay was established that uses soluble starch and 33P-labeled ATP as phosphate acceptor and donor, respectively. A constant rate of starch labeling was observed only if the ATP applied was labeled at the β position. In wild-type extracts from leaves of Arabidopsis thaliana (L.) Heynh. the maximum rate of starch phosphorylation was approximately 27 pmol min-1 (mg protein)-1. Leaf extracts from the GWD-deficient sex1 mutants of Arabidopsis showed no significant incorporation of phosphate whereas extracts from potato (Solanum tuberosum L.) tuber expressing a GWD antisense construct exhibited less activity than the wild-type control. To our knowledge this is the first time that a quantification of the starch-phosphorylating activity has been achieved in plant crude extracts.</description><subject>Adenosine Triphosphate - metabolism</subject><subject>Alleles</subject><subject>Arabidopsis - enzymology</subject><subject>Arabidopsis Proteins - metabolism</subject><subject>ATP</subject><subject>Biological and medical sciences</subject><subject>Crop harvesting</subject><subject>Enzymatic activity</subject><subject>Enzyme activity</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glucans - metabolism</subject><subject>Leaves</subject><subject>Metabolism</subject><subject>Monosaccharide Transport Proteins - metabolism</subject><subject>Phosphates</subject><subject>Phosphorus Radioisotopes</subject><subject>Phosphorylation</subject><subject>Photosynthesis, respiration. Anabolism, catabolism</subject><subject>Plant extracts</subject><subject>Plant Extracts - isolation & purification</subject><subject>Plant Extracts - metabolism</subject><subject>Plant Leaves - enzymology</subject><subject>Plant physiology and development</subject><subject>Plant tissues</subject><subject>Plants</subject><subject>Random allocation</subject><subject>Solanum tuberosum - enzymology</subject><subject>Starch</subject><subject>Starch - metabolism</subject><subject>Starch Phosphorylase - metabolism</subject><subject>Starches</subject><subject>Tubers</subject><issn>0032-0935</issn><issn>1432-2048</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpdkUtLAzEUhYMoWqs_wIUyCLobvXlMkllKfULFja5DmmbslHmZZMTx15vaYsFFCOR899x7TxA6wXCFAcS1B2AkSwFICjnFKd5BI8woSQkwuYtGAPRXyQ7QofdLgCgKsY8OMOGECS5G6PnWBuvqstGhbJukLZKwsIkP2plF2i1aH48bqqg274ltvofaJtqE8rMMQ1I2SVfpJiT2K7j46o_QXqErb4839xi93d-9Th7T6cvD0-RmmhoqeEiZ1qzIjNBUkCLjGdEEZGaF5ZpZwWQ-y5iZzWeSSZtTyrA2TBAhcxpXlSSjY3S59u1c-9FbH1RdemOrOIxte68EBSFlXH-Mzv-By7Z3TZwtMoxJwhmPEF5DxrXeO1uozpW1doPCoFZBq3XQKnZXq6AVjjVnG-N-Vtv5tmKTbAQuNoD2RleF040p_ZZjHHOIZmN0uuaWPrTuTyeUShE_i_4AIG2ObA</recordid><startdate>20030301</startdate><enddate>20030301</enddate><creator>Ritte, Gerhard</creator><creator>Steup, Martin</creator><creator>Kossmann, Jens</creator><creator>Lloyd, James R.</creator><general>Springer-Verlag</general><general>Springer</general><general>Springer Nature B.V</general><scope>IQODW</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>3V.</scope><scope>7QP</scope><scope>7QR</scope><scope>7TM</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20030301</creationdate><title>Determination of the starch-phosphorylating enzyme activity in plant extracts</title><author>Ritte, Gerhard ; Steup, Martin ; Kossmann, Jens ; Lloyd, James R.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c376t-4aa4f5c7a372f5652a2085e7e6a4e7489b54cbdb848e93341ac47278930028253</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Adenosine Triphosphate - metabolism</topic><topic>Alleles</topic><topic>Arabidopsis - enzymology</topic><topic>Arabidopsis Proteins - metabolism</topic><topic>ATP</topic><topic>Biological and medical sciences</topic><topic>Crop harvesting</topic><topic>Enzymatic activity</topic><topic>Enzyme activity</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glucans - metabolism</topic><topic>Leaves</topic><topic>Metabolism</topic><topic>Monosaccharide Transport Proteins - metabolism</topic><topic>Phosphates</topic><topic>Phosphorus Radioisotopes</topic><topic>Phosphorylation</topic><topic>Photosynthesis, respiration. Anabolism, catabolism</topic><topic>Plant extracts</topic><topic>Plant Extracts - isolation & purification</topic><topic>Plant Extracts - metabolism</topic><topic>Plant Leaves - enzymology</topic><topic>Plant physiology and development</topic><topic>Plant tissues</topic><topic>Plants</topic><topic>Random allocation</topic><topic>Solanum tuberosum - enzymology</topic><topic>Starch</topic><topic>Starch - metabolism</topic><topic>Starch Phosphorylase - metabolism</topic><topic>Starches</topic><topic>Tubers</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ritte, Gerhard</creatorcontrib><creatorcontrib>Steup, Martin</creatorcontrib><creatorcontrib>Kossmann, Jens</creatorcontrib><creatorcontrib>Lloyd, James R.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>Proquest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Planta</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ritte, Gerhard</au><au>Steup, Martin</au><au>Kossmann, Jens</au><au>Lloyd, James R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Determination of the starch-phosphorylating enzyme activity in plant extracts</atitle><jtitle>Planta</jtitle><addtitle>Planta</addtitle><date>2003-03-01</date><risdate>2003</risdate><volume>216</volume><issue>5</issue><spage>798</spage><epage>801</epage><pages>798-801</pages><issn>0032-0935</issn><eissn>1432-2048</eissn><coden>PLANAB</coden><abstract>For quantification of α-glucan, water dikinase (GWD) activity in crude extracts of plant tissues a radio-labeling assay was established that uses soluble starch and 33P-labeled ATP as phosphate acceptor and donor, respectively. 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| subjects | Adenosine Triphosphate - metabolism Alleles Arabidopsis - enzymology Arabidopsis Proteins - metabolism ATP Biological and medical sciences Crop harvesting Enzymatic activity Enzyme activity Fundamental and applied biological sciences. Psychology Glucans - metabolism Leaves Metabolism Monosaccharide Transport Proteins - metabolism Phosphates Phosphorus Radioisotopes Phosphorylation Photosynthesis, respiration. Anabolism, catabolism Plant extracts Plant Extracts - isolation & purification Plant Extracts - metabolism Plant Leaves - enzymology Plant physiology and development Plant tissues Plants Random allocation Solanum tuberosum - enzymology Starch Starch - metabolism Starch Phosphorylase - metabolism Starches Tubers |
| title | Determination of the starch-phosphorylating enzyme activity in plant extracts |
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