Method for lipidomic analysis: p53 expression modulation of sulfatide, ganglioside, and phospholipid composition of U87 MG glioblastoma cells

Research output: Contribution to journalArticle

Abstract

Lipidomics can complement genomics and proteomics by providing new insight into dynamic changes in biomembranes; however, few reports in the literature have explored, on an organism-wide scale, the functional link between nonenzymatic proteins and cellular lipids. Here, we report changes induced by adenovirus-delivered wild-type p53 gene and chemotherapy of U87 MG glioblastoma cells, a treatment known to trigger apoptosis and cell cycle arrest. We compare polar lipid changes in treated cells and control cells by use of a novel, sensitive method that employs lipid extraction, one-step liquid chromatography separation, high-resolution mass analysis, and Kendrick mass defect analysis. Nano-LC FT-ICR MS and quadrupole linear ion trap MS/MS analysis of polar lipids yields hundreds of unique assignments of glyco- and phospholipids at sub-ppm mass accuracy and high resolving power (m/Deltam50% = 200 000 at m/z 400) at 1 s/scan. MS/MS data confirm molecular structures in many instances. Sulfatides are most highly modulated by wild-type p53 treatment. The treatment also leads to an increase in phospholipids such as phosphatidyl inositols, phosphatidyl serines, phosphatidyl glycerols, and phosphatidyl ethanolamines. An increase in hydroxylated phospholipids is especially noteworthy. Also, a decrease in the longer chain gangliosides, GD1 and GM1b, is observed in wild-type p53 (treated) cells.

Details

Authors
  • Huan He
  • Charles A. Conrad
  • Carol L Nilsson
  • Yongjie Ji
  • Tanner M Schaub
  • Alan G Marshall
  • Mark R Emmett
External organisations
  • Florida State University
Research areas and keywords

Keywords

  • Cell Line, Tumor, Chromatography, Liquid, Gangliosides, Glioblastoma, Humans, Molecular Structure, Phospholipids, Spectrometry, Mass, Electrospray Ionization, Spectroscopy, Fourier Transform Infrared, Sulfoglycosphingolipids, Tumor Suppressor Protein p53, Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.
Original languageEnglish
Pages (from-to)8423-30
Number of pages8
JournalAnalytical Chemistry
Volume79
Issue number22
StatePublished - 2007 Nov 15
Peer-reviewedYes
Externally publishedYes