Mass spectrometric analysis of the glycosphingolipid-enriched microdomains of rat natural killer cells
Language English Country Germany Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Killer Cells, Natural chemistry MeSH
- Centrifugation, Density Gradient MeSH
- Cell Fractionation MeSH
- Glycosphingolipids chemistry isolation & purification MeSH
- Mass Spectrometry MeSH
- Rats MeSH
- Membrane Microdomains chemistry MeSH
- Cell Line, Tumor MeSH
- Proteomics MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Glycosphingolipids MeSH
Glycosphingolipid-enriched microdomains (GEM) are membrane entities that concentrate glycosylphosphatiolylinositol(GPI)-anchored, acylated and membrane proteins important for immune receptor signaling. Using rat leukemic cell line RNK-16 we have initiated proteomic studies of microdomains in natural killer (NK) cells. Isolated plasma membranes were treated with Brij 58, or Nonidet-P40, or sodium carbonate. Extracts were separated by sucrose density gradient centrifugation into very light membrane, medium light membrane and heavy fractions, and a complete protein profile was analyzed by tandem mass spectrometry. Up to 250 proteins were unambiguously identified in each analyzed fraction. The first study of the proteome of NK cell GEM revealed several new aspects including identification of molecules not expected to be expressed in rat NK cells (e.g., NAP-22) or associated with GEM (e.g., NKR-P1, CD45, CD2). Moreover, it provided clear data consolidating controversial views concerning the occurrence of major histcompatibility complex glycoproteins and RT6.1/CD73/CD38 complex in NK cells. Our results also identified a large number of receptors as candidates for future functional studies.
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