Novel Enzymatic Fluorometric Methods for High-throughput and Sensitive Measurements of All Major Phospholipid Classes in Cells

Abstract

Phospholipids are essential components of cell membranes in all organisms including animals, plants, fungi, and bacteria. Based on their molecular structures, phospholipids are classified into phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidic acid (PA), phosphatidylinositol (PI), phosphatidylglycerol (PG), cardiolipin (CL), sphingomyelin (SM), and so on. However, there has been no appropriate method for quantifying phsopholipid classes. Therefore, I have recently developed the enzymatic fluorometric assays to quantify all major phospholipid classes including PC, PE, PS, PA, PI, PG, CL, SM. These assays are simple, sensitive and high-throughput, but not time-consuming. These assays have been applied for the evaluation of the phos pholipid class compositions in cultured cells and their intracelluar organelles to investigate the func tions of enzymes and transporters. These enzymatic fluorometric assays will become key technologies for elucidating the biochemical and physiological roles of phospholipids in many fields of life sciences.