Quantitative Analysis of Cellular Phospholipid Metabolism by Enzymatic Fluorometric Assays

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  • Morita Shin-ya
    Department of Pharmacy, Shiga University of Medical Science Hospital

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  • 酵素蛍光定量法による細胞内リン脂質代謝機能解析
  • コウソ ケイコウ テイリョウホウ ニ ヨル サイボウ ナイ リン シシツ タイシャ キノウ カイセキ

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Description

Phospholipids are major components of cell membranes and plasma lipoproteins. In addition to structural roles in membranes, phospholipids are involved in many cellular processes, including cell signaling, membrane trafficking, cell growth, differentiation, migration, apoptosis, and autophagy. Phospholipids are divided into two groups, glycerophospholipids and sphingophospholipids. By the structures of head groups, glycerophospholipids are further classified into classes, such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidic acid (PA), phosphatidylinositol (PI), phosphatidylglycerol (PG) and cardiolipin (CL). Sphingomyelin (SM) is a major sphingophospholipid. However, it has been difficult to determine the cellular amount of each phospholipid class. In recent years, we have developed novel fluorometric methods for measuring PC, PE, PS, PA, SM, PG and CL using combinations of specific enzymes and Amplex Red. These new assays can provide simple, sensitive and high–throughput quantifications. By using these assays, we investigated cellular phospholipid metabolism involving cell density, PE N–methyltransferase, PS synthase 1, diacylglycerol kinase, tumor necrosis factorα, and phosphatidylglycerophosphate synthase 1. These assays for measuring phospholipid classes will be useful to clarify the cellular and physiological processes in any organisms, including animals, plants, fungi and bacteria.

Journal

  • MEMBRANE

    MEMBRANE 41 (5), 202-208, 2016

    THE MEMBRANE SOCIETY OF JAPAN

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