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| pubmed-article:10585749 | lifeskim:mentions | umls-concept:C1527178 | lld:lifeskim |
| pubmed-article:10585749 | pubmed:issue | 1 | lld:pubmed |
| pubmed-article:10585749 | pubmed:dateCreated | 2000-2-14 | lld:pubmed |
| pubmed-article:10585749 | pubmed:abstractText | A highly sensitive method for determination of insulin receptor (IR) kinase activity in whole cells, which is based on a STAT5 (signal transducer and activator of transcription 5)-dependent reporter gene assay, has been developed. We show in Rat1 fibroblasts stably overexpressing the human IR (Rat1-HIR-cl5) an insulin-dependent direct association and phosphorylation of STAT5b by IR kinase. Rat1-HIR cells transfected with a luciferase gene reporter construct under control of a STAT5-inducible promoter showed insulin-mediated induction of STAT5-dependent luciferase activity, with peak activities around 8 h of insulin treatment over a wide dose range. Transient STAT5b but not STAT5a cotransfection significantly enhanced reporter gene activity, yielding up to a fivefold induction. Addition of the IR kinase inhibitor tyrphostin AG1024 down-regulated luciferase induction in a dose-dependent manner. This is the first assay allowing determination of IR kinase activity in intact cells in a 24-well culture and a microtiter format. Kinetics of this cellular response, sensitivity range, and signal amplitude make it well suited for automation and offer the potential for establishing high-throughput screening systems for both insulin mimetic substances and IR kinase antagonists in a simple nonradioactive assay. | lld:pubmed |
| pubmed-article:10585749 | pubmed:language | eng | lld:pubmed |
| pubmed-article:10585749 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:10585749 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:10585749 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
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| pubmed-article:10585749 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:10585749 | pubmed:month | Dec | lld:pubmed |
| pubmed-article:10585749 | pubmed:issn | 0003-2697 | lld:pubmed |
| pubmed-article:10585749 | pubmed:author | pubmed-author:StormOO | lld:pubmed |
| pubmed-article:10585749 | pubmed:author | pubmed-author:MüllerGG | lld:pubmed |
| pubmed-article:10585749 | pubmed:author | pubmed-author:PfizenmaierKK | lld:pubmed |
| pubmed-article:10585749 | pubmed:author | pubmed-author:GronerBB | lld:pubmed |
| pubmed-article:10585749 | pubmed:author | pubmed-author:DöpplerHH | lld:pubmed |
| pubmed-article:10585749 | pubmed:author | pubmed-author:Horn-MüllerJJ | lld:pubmed |
| pubmed-article:10585749 | pubmed:copyrightInfo | Copyright 1999 Academic Press. | lld:pubmed |
| pubmed-article:10585749 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:10585749 | pubmed:day | 1 | lld:pubmed |
| pubmed-article:10585749 | pubmed:volume | 276 | lld:pubmed |
| pubmed-article:10585749 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:10585749 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:10585749 | pubmed:pagination | 97-104 | lld:pubmed |
| pubmed-article:10585749 | pubmed:dateRevised | 2011-11-17 | lld:pubmed |
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| pubmed-article:10585749 | pubmed:year | 1999 | lld:pubmed |
| pubmed-article:10585749 | pubmed:articleTitle | A cellular reporter assay to monitor insulin receptor kinase activity based on STAT 5-dependent luciferase gene expression. | lld:pubmed |
| pubmed-article:10585749 | pubmed:affiliation | Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, D-70569, Germany. | lld:pubmed |
| pubmed-article:10585749 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:10585749 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
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