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Protocols for Enzymatic Fluorometric Assays to Quantify Phospholipid Classes

Phospholipids, consisting of a hydrophilic head group and two hydrophobic acyl chains, are essential for the structures of cell membranes, plasma lipoproteins, biliary mixed micelles, pulmonary surfactants, and extracellular vesicles. Beyond their structural roles, phospholipids have important roles...

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Autores principales: Morita, Shin-ya, Tsuji, Tokuji, Terada, Tomohiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7037927/
https://www.ncbi.nlm.nih.gov/pubmed/32033167
http://dx.doi.org/10.3390/ijms21031032
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author Morita, Shin-ya
Tsuji, Tokuji
Terada, Tomohiro
author_facet Morita, Shin-ya
Tsuji, Tokuji
Terada, Tomohiro
author_sort Morita, Shin-ya
collection PubMed
description Phospholipids, consisting of a hydrophilic head group and two hydrophobic acyl chains, are essential for the structures of cell membranes, plasma lipoproteins, biliary mixed micelles, pulmonary surfactants, and extracellular vesicles. Beyond their structural roles, phospholipids have important roles in numerous biological processes. Thus, abnormalities in the metabolism and transport of phospholipids are involved in many diseases, including dyslipidemia, atherosclerosis, cholestasis, drug-induced liver injury, neurological diseases, autoimmune diseases, respiratory diseases, myopathies, and cancers. To further clarify the physiological, pathological, and molecular mechanisms and to identify disease biomarkers, we have recently developed enzymatic fluorometric assays for quantifying all major phospholipid classes, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidic acid, phosphatidylinositol, phosphatidylglycerol + cardiolipin, and sphingomyelin. These assays are specific, sensitive, simple, and high-throughput, and will be applicable to cells, intracellular organelles, tissues, fluids, lipoproteins, and extracellular vesicles. In this review, we present the detailed protocols for the enzymatic fluorometric measurements of phospholipid classes in cultured cells.
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spelling pubmed-70379272020-03-10 Protocols for Enzymatic Fluorometric Assays to Quantify Phospholipid Classes Morita, Shin-ya Tsuji, Tokuji Terada, Tomohiro Int J Mol Sci Review Phospholipids, consisting of a hydrophilic head group and two hydrophobic acyl chains, are essential for the structures of cell membranes, plasma lipoproteins, biliary mixed micelles, pulmonary surfactants, and extracellular vesicles. Beyond their structural roles, phospholipids have important roles in numerous biological processes. Thus, abnormalities in the metabolism and transport of phospholipids are involved in many diseases, including dyslipidemia, atherosclerosis, cholestasis, drug-induced liver injury, neurological diseases, autoimmune diseases, respiratory diseases, myopathies, and cancers. To further clarify the physiological, pathological, and molecular mechanisms and to identify disease biomarkers, we have recently developed enzymatic fluorometric assays for quantifying all major phospholipid classes, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidic acid, phosphatidylinositol, phosphatidylglycerol + cardiolipin, and sphingomyelin. These assays are specific, sensitive, simple, and high-throughput, and will be applicable to cells, intracellular organelles, tissues, fluids, lipoproteins, and extracellular vesicles. In this review, we present the detailed protocols for the enzymatic fluorometric measurements of phospholipid classes in cultured cells. MDPI 2020-02-04 /pmc/articles/PMC7037927/ /pubmed/32033167 http://dx.doi.org/10.3390/ijms21031032 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Review
Morita, Shin-ya
Tsuji, Tokuji
Terada, Tomohiro
Protocols for Enzymatic Fluorometric Assays to Quantify Phospholipid Classes
title Protocols for Enzymatic Fluorometric Assays to Quantify Phospholipid Classes
title_full Protocols for Enzymatic Fluorometric Assays to Quantify Phospholipid Classes
title_fullStr Protocols for Enzymatic Fluorometric Assays to Quantify Phospholipid Classes
title_full_unstemmed Protocols for Enzymatic Fluorometric Assays to Quantify Phospholipid Classes
title_short Protocols for Enzymatic Fluorometric Assays to Quantify Phospholipid Classes
title_sort protocols for enzymatic fluorometric assays to quantify phospholipid classes
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7037927/
https://www.ncbi.nlm.nih.gov/pubmed/32033167
http://dx.doi.org/10.3390/ijms21031032
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