Kouzel, Ivan U. ; Pohlentz, Gottfried; Schmitz, Julia S.; Steil, Daniel ; Humpf, Hans-Ulrich ; Karch, Helge; Müthing, Johannes: Shiga Toxin Glycosphingolipid Receptors in Human Caco-2 and HCT-8 Colon Epithelial Cell Lines
Inhalt
- Results
- Identification of Neutral GSLs of the Globo-Series in Caco-2 and HCT-8 Cells
- Structural Characterization of Sphingolipids from the Neutral GSL Fraction of Caco-2 and HCT-8 Cells
- Structural Characterization of GlcCer and GalCer Lipoforms Detected in the Neutral GSL Fraction of Caco-2 and HCT-8 Cells
- Relative Content of the Various Gb3Cer and Gb4Cer Lipoforms of Caco-2 and HCT-8 Cells
- Identification and Structural Characterization of Stx2a-Binding GSLs
- Distribution of Gb3Cer and Gb4Cer to DRM and NonDRM Fractions
- Stx2a-Mediated Cellular Damage of Caco-2 and HCT-8 Cells
- Discussion
- Conclusions
- Materials and Methods
- Cultivation of Caco-2, HCT-8 and Vero-B4 Cell Lines and Stx2a Cytotoxicity Assay
- Isolation of Lipids and Purification of GSLs from Cultured Cell Lines
- Preparation of Sucrose Density Gradient Fractions by Ultracentrifugation
- Isolation of Lipids from Sucrose Density Gradient Fractions
- Lipid References, Antibodies and Stx2a
- Thin-Layer Chromatography and Lipid Staining
- TLC Overlay Assay and Lipid Semiquantification
- Mass Spectrometry of GSLs