학술논문

Analysis of the Phosphoinositide Composition of Subcellular Membrane Fractions
Document Type
Chapter
Author
Source
Lipid Signaling ProtocolsLipid Signaling Protocols. 01/01/2016. 1376:213-227
Subject
Phosphoinositide
Subcellular fractionation
Metabolic label ing
Organelle
Life Sciences
Cell Biology
Membrane Biology
Lipidology
Biomedical and Life Sciences
Language
English
ISSN
1940-6029
1064-3745
Abstract
Phosphoinositides play critical roles in the transduction of extracellular signals through the plasma membrane and also in endomembrane events important for vesicle trafficking and organelle function (Di Paolo and De Camilli, Nature 443(7112):651–657, 2006). The response triggered by these lipids is heavily dependent on the microenvironment in which they are found. HPLC analysis of labeled phosphoinositides allows quantification of the levels of each phosphoinositide species relative to their precursor, phosphatidylinositol. When combined with subcellular fractionation techniques, this strategy allows measurement of the relative phosphoinositide composition of each membrane fraction or organelle and determination of the microenvironment in which each species is enriched. Here, we describe the steps to separate and quantify total or localized phosphoinositides from cultured cells.