Quantitative Analysis of Phospholipids in Functionally Important Membrane Domains from RBL-2H3 Mast Cells Using Tandem High-Resolution Mass Spectrometry

作者：Einar K. Fridriksson, Petia A. Shipkova, Erin D. Sheets, David Holowka, Barbara A. Baird, Fred W. McLafferty · 发表于：Biochemistry · 年份：1999 · DOI：10.1021/bi9828324 · 被引用次数：287 · 研究领域：Lipid Membrane Structure and Behavior、Mass Spectrometry Techniques and Applications、Ion channel regulation and function

We recently showed that ligand-mediated cross-linking of FcepsilonRI, the high-affinity receptor for immunoglobulin E, on RBL-2H3 mast cells results in its co-isolation with detergent-resistant membranes (DRM) and its consequent tyrosine phosphorylation by the co-localized tyrosine kinase Lyn that is a critical early event in signaling by this receptor [Field et al. (1997) J. Biol. Chem. 272, 4276-4280]. As part of efforts to determine the structural bases for these interactions, we examined the phospholipid composition of DRM vesicles isolated from RBL-2H3 cells under conditions that preserve FcepsilonRI association. We used positive and negative mode electrospray Fourier transform ion cyclotron resonance mass spectrometry to compare quantitatively the phospholipid composition of isolated DRM to that of total cell lipids and to a plasma membrane preparation. From these analyses, over 90 different phospholipid species were spectrally resolved and unambiguously identified; more than two-thirds of these were determined with a precision of +/-0.5% (absolute) or less. Quantitative characterization of lipid profiles shows that isolated DRM are substantially enriched in sphingomyelin and in glycerophospholipids with a higher degree of saturation as compared to total cellular lipids. Plasma membrane vesicles isolated from RBL-2H3 cells by chemically induced blebbing exhibit a degree of phospholipid saturation that is intermediate between DRM and total cellular lipids, and significan...