2018 Fiscal Year Final Research Report
Elucidation of disease mechanisms and search for biomarkers by using enzymatic fluorometric assays for quantifying all phospholipids
| Project/Area Number |
15H05660
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| Research Category |
Grant-in-Aid for Young Scientists (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | Shiga University of Medical Science |
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Principal Investigator |
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| Research Collaborator |
Tsuji Tokuji
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| Project Period (FY) |
2015-04-01 – 2019-03-31
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| Keywords | リン脂質 / 酵素蛍光定量法 / ホスファチジルイノシトール / ホスファチジルグリセロール / カルジオリピン / 細胞密度 |
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| Outline of Final Research Achievements |
Phospholipids are the main constituents of cell membranes and plasma lipoproteins. Based on structures, phospholipids are classified into classes such as phosphatidylcholine (PC) and phosphatidylinositol (PI). However, there has been no highly sensitive and high-throughput method for measuring phospholipids. In this study, in addition to our previously developed assays, we developed the enzymatic fluorometric assays for quantifying all major phospholipid classes. Furthermore, by using these enzymatic fluorometric assays for measurements of all major phospholipid classes, we evaluated in detail the effects of cell density of enzyme expression on phospholipid compositions in cultured cells. We also elucidated the mechanism of biliary phospholipid secretion.
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| Free Research Field |
脂質生化学
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| Academic Significance and Societal Importance of the Research Achievements |
従来、リン脂質の定量は、TLC/リン定量法などにより行われてきたが、微量の生体試料に含まれているリン脂質をクラスごとに定量解析するには十分でなかった。そこで、これまでに研究代表者はリン脂質クラスのうちPA・PC・PE・PS・SMに対する酵素蛍光定量法を確立していた。本研究では、PIとPG・CLの酵素蛍光定量法の開発に成功した。これにより、主要な全リン脂質クラスに対する高感度・高精度かつハイスループットな網羅的一斉定量法がついに完成した。本定量法は、培養細胞実験に加えて動物実験や植物・微生物実験を含む生命科学研究全般ならびにバイオマーカー探索等の臨床研究に活用されることが期待できる。
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