Development of simple method for detecting the sphingolipid by using click chmistry

2014 Fiscal Year Final Research Report

• Project/Area Number: 25670024
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biological pharmacy
• Research Institution: Osaka University
• Principal Investigator: NISHI TSUYOSHI 大阪大学, 産業科学研究所, 准教授 (60403002)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: S1P / 輸送体 / クリックケミストリー / SPNS2 / 蛍光プローブ

Outline of Final Research Achievements

S1P transporter should be a good candidate for development of immunosuppressive drugs without severe side effect. However, quantification methods of S1P are time consuming or expensive for high throughput screening. We tried to develop the simple assay method using fluorescent-labeled S1P. The fluorescent-labeled S1P was secreted from various culture cells and was not a substrate of SPNS2.Therefore,to develop the detection system for SPNS2 activity,we tried to synthesize alkyn-labeled sphingosine. We success to synthesize alkyn-labeled sphingosine and fluorescent-labeled azide. These compound was showed crick reaction in optimized condition and fluorescent-labeled sphingosine was detected in μM level. To perform the high throughput screening of SPNS2 inhibitors, further development is necesarry to adapt this method for detecting the alkyn-S1P secreted from the cells.

Free Research Field

生化学