Full metadata record
DC Field | Value | Language |
---|---|---|
dc.contributor.author | 오세관 | * |
dc.date.accessioned | 2016-08-28T12:08:33Z | - |
dc.date.available | 2016-08-28T12:08:33Z | - |
dc.date.issued | 2006 | * |
dc.identifier.issn | 0253-6269 | * |
dc.identifier.other | OAK-3679 | * |
dc.identifier.uri | https://dspace.ewha.ac.kr/handle/2015.oak/219782 | - |
dc.description.abstract | Activation of Sphingosine kinase (Sphk) increases a bioactive lipid, sphingosine 1-phosphate (S1P) and has been observed in a variety of cancer cells. Therefore, inhibition of Sphk activity was an important target for the development of anticancer drugs. As a searching tool for Sphk inhibitor, we developed fluorescent Sphk activity assay combined with high performance liquid chromatography (HPLC). Previously we established murine teraticarcinoma mutant F9-12 cells which lack S1P lyase and stably express Sphk1. By using F9-12 cells, optimal assay conditions were established as follows; 100 μM of C 17-Sph and 30 μg protein of F9-12 cells lysate in 20 min. Sphingosine analog C17-Sph was efficiently phosphorylated by Sphk activity (Km :67.08 μM, Vmax :1507.5 pmol/min/mg). New product C17-S1P was separated from S1P in reversed-phase HPLC. In optimized conditions, 300 nM of phorbol 12-myristate 13-acetate (PMA) increased Sphk activity approximately twice while 20 μM of N,N-dimethylsphingosine (DMS) reduced 70% of Sphk activity in F9-12 cells lysate. In conclusion, we established non-radioactive but convenient Sphk assay system by using HPLC and F9-12 cells. | * |
dc.language | English | * |
dc.title | Sphingosine kinase assay system with fluorescent detection in high performance liquid chromatography | * |
dc.type | Article | * |
dc.relation.issue | 11 | * |
dc.relation.volume | 29 | * |
dc.relation.index | SCIE | * |
dc.relation.index | SCOPUS | * |
dc.relation.index | KCI | * |
dc.relation.startpage | 1049 | * |
dc.relation.lastpage | 1054 | * |
dc.relation.journaltitle | Archives of Pharmacal Research | * |
dc.identifier.wosid | WOS:000242385600017 | * |
dc.identifier.scopusid | 2-s2.0-33845356305 | * |
dc.author.google | Jin Y.-X. | * |
dc.author.google | Yoo H.-S. | * |
dc.author.google | Kihara A. | * |
dc.author.google | Choi C.-H. | * |
dc.author.google | Oh S. | * |
dc.author.google | Moon D.-C. | * |
dc.author.google | Igarashi Y. | * |
dc.author.google | Lee Y.-M. | * |
dc.contributor.scopusid | 오세관(7404103757) | * |
dc.date.modifydate | 20240118133340 | * |