An ultrasensitive competitive binding assay for the detection of toxins affecting protein phosphatases
An ultrasensitive assay is described for microcystin-LR and other substances (microcystins, nodularin, okadaic acid, calyculin A, tautomycin) which block the active site of protein phosphatases (PP) 1 and 2A. The assay is based on competition between the unknown sample and [ 125I]microcystin-YR for...
Gespeichert in:
| Veröffentlicht in: | Toxicon (Oxford) 2000-03, Vol.38 (3), p.347-360 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 360 |
|---|---|
| container_issue | 3 |
| container_start_page | 347 |
| container_title | Toxicon (Oxford) |
| container_volume | 38 |
| creator | Serres, M.H. Fladmark, K.E. Døskeland, S.O. |
| description | An ultrasensitive assay is described for microcystin-LR and other substances (microcystins, nodularin, okadaic acid, calyculin A, tautomycin) which block the active site of protein phosphatases (PP) 1 and 2A. The assay is based on competition between the unknown sample and [
125I]microcystin-YR for binding to the catalytic subunit of PP2A. The PP2A-bound [
125I]microcystin-YR was stable (half-time of dissociation=1.8 h), allowing non-bound [
125I]microcystin-YR to be removed by Sephadex G-50 size-exclusion chromatography. Compared to current assays based on inhibition of protein phosphatase activity, the present assay was more robust against interference (from fluoride, ATP, histone, and casein), and had an even better sensitivity. The detection limit was below 50 pM (2.5 fmol) for nodularin and microcystin-LR, and below 200 pM (10 fmol) for okadaic acid. The method was used successfully to detect extremely low concentrations of either microcystin or nodularin in drinking water or seawater, and okadaic acid in shellfish extract. |
| doi_str_mv | 10.1016/S0041-0101(99)00163-4 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_17442270</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0041010199001634</els_id><sourcerecordid>17442270</sourcerecordid><originalsourceid>FETCH-LOGICAL-c421t-8627e3d49d97c866bd1bd8ab9a22330b33d36c4a568fec3793badb3ffaad30ef3</originalsourceid><addsrcrecordid>eNqFkE2LFDEQhoMo7uzqT1ByENFDa76mu3OSZXFVWPCgnkM6qTiRnqRNZRb335vZHtSbpyqKp6peHkKecfaGM96__cKY4h1r_SutX7M2kp16QDZ8HHQn-ZY9JJs_yBk5R_zBGJOj7h-TM876XjOhNiRcJnqYa7EICWONt0Bd3i9Q136Kycf0nVpEe0dDLrTugHqo4GrMieZAa_4VE1IbwnHW2KXkCjHRZZdx2dnaTuMT8ijYGeHpqV6Qb9fvv1597G4-f_h0dXnTOSV47cZeDCC90l4Pbuz7yfPJj3bSVggp2SSll71TdtuP7ZsctJysn2QI1nrJIMgL8nK920L8PABWs4_oYJ5tgnxAwwelhBhYA7cr6EpGLBDMUuLeljvDmTkKNveCzdGe0drcCzaq7T0_PThMe_D_bK1GG_DiBFh0dg7FJhfxLydGIbeiYe9WDJqN2wjFoIuQHPhYmkfjc_xPkt8YApnc</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>17442270</pqid></control><display><type>article</type><title>An ultrasensitive competitive binding assay for the detection of toxins affecting protein phosphatases</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Serres, M.H. ; Fladmark, K.E. ; Døskeland, S.O.</creator><creatorcontrib>Serres, M.H. ; Fladmark, K.E. ; Døskeland, S.O.</creatorcontrib><description>An ultrasensitive assay is described for microcystin-LR and other substances (microcystins, nodularin, okadaic acid, calyculin A, tautomycin) which block the active site of protein phosphatases (PP) 1 and 2A. The assay is based on competition between the unknown sample and [
125I]microcystin-YR for binding to the catalytic subunit of PP2A. The PP2A-bound [
125I]microcystin-YR was stable (half-time of dissociation=1.8 h), allowing non-bound [
125I]microcystin-YR to be removed by Sephadex G-50 size-exclusion chromatography. Compared to current assays based on inhibition of protein phosphatase activity, the present assay was more robust against interference (from fluoride, ATP, histone, and casein), and had an even better sensitivity. The detection limit was below 50 pM (2.5 fmol) for nodularin and microcystin-LR, and below 200 pM (10 fmol) for okadaic acid. The method was used successfully to detect extremely low concentrations of either microcystin or nodularin in drinking water or seawater, and okadaic acid in shellfish extract.</description><identifier>ISSN: 0041-0101</identifier><identifier>EISSN: 1879-3150</identifier><identifier>DOI: 10.1016/S0041-0101(99)00163-4</identifier><identifier>PMID: 10669024</identifier><identifier>CODEN: TOXIA6</identifier><language>eng</language><publisher>Oxford: Elsevier Ltd</publisher><subject>Bacterial Toxins - analysis ; Bacterial Toxins - chemistry ; Bacterial Toxins - pharmacology ; Bacteriology ; Binding Sites - drug effects ; Binding, Competitive - drug effects ; Biological and medical sciences ; Biological Assay ; Fundamental and applied biological sciences. Psychology ; Iodine - chemistry ; Microbiology ; Microcystins ; nodularin ; Okadaic Acid - chemistry ; Okadaic Acid - pharmacology ; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains ; Peptides, Cyclic - analysis ; Peptides, Cyclic - chemistry ; Peptides, Cyclic - pharmacology ; phosphatase ; Phosphoprotein Phosphatases - antagonists & inhibitors ; Phosphoprotein Phosphatases - metabolism ; Seawater - analysis ; Shellfish - analysis ; Time Factors ; Tissue Extracts - analysis ; Toxins, Biological - analysis ; Toxins, Biological - chemistry ; Toxins, Biological - pharmacology ; Water Supply - analysis</subject><ispartof>Toxicon (Oxford), 2000-03, Vol.38 (3), p.347-360</ispartof><rights>1999 Elsevier Science Ltd</rights><rights>2000 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c421t-8627e3d49d97c866bd1bd8ab9a22330b33d36c4a568fec3793badb3ffaad30ef3</citedby><cites>FETCH-LOGICAL-c421t-8627e3d49d97c866bd1bd8ab9a22330b33d36c4a568fec3793badb3ffaad30ef3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0041-0101(99)00163-4$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,781,785,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1282352$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10669024$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Serres, M.H.</creatorcontrib><creatorcontrib>Fladmark, K.E.</creatorcontrib><creatorcontrib>Døskeland, S.O.</creatorcontrib><title>An ultrasensitive competitive binding assay for the detection of toxins affecting protein phosphatases</title><title>Toxicon (Oxford)</title><addtitle>Toxicon</addtitle><description>An ultrasensitive assay is described for microcystin-LR and other substances (microcystins, nodularin, okadaic acid, calyculin A, tautomycin) which block the active site of protein phosphatases (PP) 1 and 2A. The assay is based on competition between the unknown sample and [
125I]microcystin-YR for binding to the catalytic subunit of PP2A. The PP2A-bound [
125I]microcystin-YR was stable (half-time of dissociation=1.8 h), allowing non-bound [
125I]microcystin-YR to be removed by Sephadex G-50 size-exclusion chromatography. Compared to current assays based on inhibition of protein phosphatase activity, the present assay was more robust against interference (from fluoride, ATP, histone, and casein), and had an even better sensitivity. The detection limit was below 50 pM (2.5 fmol) for nodularin and microcystin-LR, and below 200 pM (10 fmol) for okadaic acid. The method was used successfully to detect extremely low concentrations of either microcystin or nodularin in drinking water or seawater, and okadaic acid in shellfish extract.</description><subject>Bacterial Toxins - analysis</subject><subject>Bacterial Toxins - chemistry</subject><subject>Bacterial Toxins - pharmacology</subject><subject>Bacteriology</subject><subject>Binding Sites - drug effects</subject><subject>Binding, Competitive - drug effects</subject><subject>Biological and medical sciences</subject><subject>Biological Assay</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Iodine - chemistry</subject><subject>Microbiology</subject><subject>Microcystins</subject><subject>nodularin</subject><subject>Okadaic Acid - chemistry</subject><subject>Okadaic Acid - pharmacology</subject><subject>Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains</subject><subject>Peptides, Cyclic - analysis</subject><subject>Peptides, Cyclic - chemistry</subject><subject>Peptides, Cyclic - pharmacology</subject><subject>phosphatase</subject><subject>Phosphoprotein Phosphatases - antagonists & inhibitors</subject><subject>Phosphoprotein Phosphatases - metabolism</subject><subject>Seawater - analysis</subject><subject>Shellfish - analysis</subject><subject>Time Factors</subject><subject>Tissue Extracts - analysis</subject><subject>Toxins, Biological - analysis</subject><subject>Toxins, Biological - chemistry</subject><subject>Toxins, Biological - pharmacology</subject><subject>Water Supply - analysis</subject><issn>0041-0101</issn><issn>1879-3150</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE2LFDEQhoMo7uzqT1ByENFDa76mu3OSZXFVWPCgnkM6qTiRnqRNZRb335vZHtSbpyqKp6peHkKecfaGM96__cKY4h1r_SutX7M2kp16QDZ8HHQn-ZY9JJs_yBk5R_zBGJOj7h-TM876XjOhNiRcJnqYa7EICWONt0Bd3i9Q136Kycf0nVpEe0dDLrTugHqo4GrMieZAa_4VE1IbwnHW2KXkCjHRZZdx2dnaTuMT8ijYGeHpqV6Qb9fvv1597G4-f_h0dXnTOSV47cZeDCC90l4Pbuz7yfPJj3bSVggp2SSll71TdtuP7ZsctJysn2QI1nrJIMgL8nK920L8PABWs4_oYJ5tgnxAwwelhBhYA7cr6EpGLBDMUuLeljvDmTkKNveCzdGe0drcCzaq7T0_PThMe_D_bK1GG_DiBFh0dg7FJhfxLydGIbeiYe9WDJqN2wjFoIuQHPhYmkfjc_xPkt8YApnc</recordid><startdate>20000301</startdate><enddate>20000301</enddate><creator>Serres, M.H.</creator><creator>Fladmark, K.E.</creator><creator>Døskeland, S.O.</creator><general>Elsevier Ltd</general><general>Elsevier Science</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>7U7</scope><scope>C1K</scope><scope>F1W</scope><scope>H95</scope><scope>H97</scope><scope>L.G</scope></search><sort><creationdate>20000301</creationdate><title>An ultrasensitive competitive binding assay for the detection of toxins affecting protein phosphatases</title><author>Serres, M.H. ; Fladmark, K.E. ; Døskeland, S.O.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c421t-8627e3d49d97c866bd1bd8ab9a22330b33d36c4a568fec3793badb3ffaad30ef3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Bacterial Toxins - analysis</topic><topic>Bacterial Toxins - chemistry</topic><topic>Bacterial Toxins - pharmacology</topic><topic>Bacteriology</topic><topic>Binding Sites - drug effects</topic><topic>Binding, Competitive - drug effects</topic><topic>Biological and medical sciences</topic><topic>Biological Assay</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Iodine - chemistry</topic><topic>Microbiology</topic><topic>Microcystins</topic><topic>nodularin</topic><topic>Okadaic Acid - chemistry</topic><topic>Okadaic Acid - pharmacology</topic><topic>Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains</topic><topic>Peptides, Cyclic - analysis</topic><topic>Peptides, Cyclic - chemistry</topic><topic>Peptides, Cyclic - pharmacology</topic><topic>phosphatase</topic><topic>Phosphoprotein Phosphatases - antagonists & inhibitors</topic><topic>Phosphoprotein Phosphatases - metabolism</topic><topic>Seawater - analysis</topic><topic>Shellfish - analysis</topic><topic>Time Factors</topic><topic>Tissue Extracts - analysis</topic><topic>Toxins, Biological - analysis</topic><topic>Toxins, Biological - chemistry</topic><topic>Toxins, Biological - pharmacology</topic><topic>Water Supply - analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Serres, M.H.</creatorcontrib><creatorcontrib>Fladmark, K.E.</creatorcontrib><creatorcontrib>Døskeland, S.O.</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>Toxicology Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><jtitle>Toxicon (Oxford)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Serres, M.H.</au><au>Fladmark, K.E.</au><au>Døskeland, S.O.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An ultrasensitive competitive binding assay for the detection of toxins affecting protein phosphatases</atitle><jtitle>Toxicon (Oxford)</jtitle><addtitle>Toxicon</addtitle><date>2000-03-01</date><risdate>2000</risdate><volume>38</volume><issue>3</issue><spage>347</spage><epage>360</epage><pages>347-360</pages><issn>0041-0101</issn><eissn>1879-3150</eissn><coden>TOXIA6</coden><abstract>An ultrasensitive assay is described for microcystin-LR and other substances (microcystins, nodularin, okadaic acid, calyculin A, tautomycin) which block the active site of protein phosphatases (PP) 1 and 2A. The assay is based on competition between the unknown sample and [
125I]microcystin-YR for binding to the catalytic subunit of PP2A. The PP2A-bound [
125I]microcystin-YR was stable (half-time of dissociation=1.8 h), allowing non-bound [
125I]microcystin-YR to be removed by Sephadex G-50 size-exclusion chromatography. Compared to current assays based on inhibition of protein phosphatase activity, the present assay was more robust against interference (from fluoride, ATP, histone, and casein), and had an even better sensitivity. The detection limit was below 50 pM (2.5 fmol) for nodularin and microcystin-LR, and below 200 pM (10 fmol) for okadaic acid. The method was used successfully to detect extremely low concentrations of either microcystin or nodularin in drinking water or seawater, and okadaic acid in shellfish extract.</abstract><cop>Oxford</cop><pub>Elsevier Ltd</pub><pmid>10669024</pmid><doi>10.1016/S0041-0101(99)00163-4</doi><tpages>14</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0041-0101 |
| ispartof | Toxicon (Oxford), 2000-03, Vol.38 (3), p.347-360 |
| issn | 0041-0101 1879-3150 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_17442270 |
| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Bacterial Toxins - analysis Bacterial Toxins - chemistry Bacterial Toxins - pharmacology Bacteriology Binding Sites - drug effects Binding, Competitive - drug effects Biological and medical sciences Biological Assay Fundamental and applied biological sciences. Psychology Iodine - chemistry Microbiology Microcystins nodularin Okadaic Acid - chemistry Okadaic Acid - pharmacology Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains Peptides, Cyclic - analysis Peptides, Cyclic - chemistry Peptides, Cyclic - pharmacology phosphatase Phosphoprotein Phosphatases - antagonists & inhibitors Phosphoprotein Phosphatases - metabolism Seawater - analysis Shellfish - analysis Time Factors Tissue Extracts - analysis Toxins, Biological - analysis Toxins, Biological - chemistry Toxins, Biological - pharmacology Water Supply - analysis |
| title | An ultrasensitive competitive binding assay for the detection of toxins affecting protein phosphatases |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-13T11%3A45%3A49IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=An%20ultrasensitive%20competitive%20binding%20assay%20for%20the%20detection%20of%20toxins%20affecting%20protein%20phosphatases&rft.jtitle=Toxicon%20(Oxford)&rft.au=Serres,%20M.H.&rft.date=2000-03-01&rft.volume=38&rft.issue=3&rft.spage=347&rft.epage=360&rft.pages=347-360&rft.issn=0041-0101&rft.eissn=1879-3150&rft.coden=TOXIA6&rft_id=info:doi/10.1016/S0041-0101(99)00163-4&rft_dat=%3Cproquest_cross%3E17442270%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=17442270&rft_id=info:pmid/10669024&rft_els_id=S0041010199001634&rfr_iscdi=true |