Identification of phosphatidylcholine-specifc phospholipase C

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K14470
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Functional biochemistry
• Research Institution: Chiba University
• Principal Investigator: Sakane Fumio 千葉大学, 大学院理学研究科, 教授 (10183815)
• Co-Investigator(Renkei-kenkyūsha): SAKAI HIROMICHI 島根大学, 総合科学研究支援センター, 助教 (00375255)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: ホスファチジルコリン-特異的ホスホリパーゼC / ジアシルグリセロールキナーゼ / 生体膜 / 動脈硬化

Outline of Final Research Achievements

Although the activity of mammalian phosphatidylcholine (PC)-specific phospholipase C (PC-PLC) was discovered approximately 30 years ago, neither the protein nor its gene has been identified. Because we recently revealed that diacylglycerol kinase (DGK) delta bound to PC-PLC, we attempted to identify PC-PLC using the interaction. In the present study, we found that the coiled-coil region in DGKdelta interacted with PC-PLC. Moreover, we established a new LC-MS-based PC-PLC assay, which has better analytical parameters, such as a lower background (0.02% versus 94%), higher S/B (4242 versus 1.06), and lower limit of quantitation (0.04 pmol versus 0.69 pmol of PC-PLC), than the conventional, indirect fluorometric assay.

Free Research Field

生化学