Protein and lipid analysis of detergent-resistant membranes isolated from bovine kidney
| Autor: | Michel Lagarde, Annie-France Prigent, Michel Becchi, Françoise Besson, Bernard Roux, Stéphanie Bonnin, Madeleine Dubois, Karim El Kirat, Christophe Grangeasse, Claire Giraud |
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| Přispěvatelé: | Deleage, Gilbert |
| Rok vydání: | 2003 |
| Předmět: |
Dipeptidases
Detergents GPI-Linked Proteins Kidney Biochemistry Palmitic acid Membrane Lipids chemistry.chemical_compound Membrane Microdomains Phosphatidylcholine Spectroscopy Fourier Transform Infrared [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology Animals Phospholipids chemistry.chemical_classification Phosphatidylethanolamine Chromatography Fatty Acids Membrane Proteins Fatty acid General Medicine Alkaline Phosphatase Membrane chemistry Saturated fatty acid Alkaline phosphatase Cattle lipids (amino acids peptides and proteins) Sphingomyelin Protein Kinases |
| Zdroj: | Biochimie. 85:1237-1244 |
| ISSN: | 0300-9084 |
| DOI: | 10.1016/j.biochi.2003.11.004 |
| Popis: | Detergent-resistant membranes (DRM) were prepared from bovine kidney cortex. The criterion used to test their purification was the increase in the activity of a GPI membrane-anchored protein, the alkaline phosphatase. Its association with specific proteins and lipids was tested. Two successive Triton X-100 treatments followed by purification on sucrose gradient at 4 °C were necessary to obtain DRM with a maximum of alkaline phosphatase activity and a typical protein pattern. A third Triton treatment did not alter this DRM composition. Among the enriched protein, we identified, by mass spectrometry, a microsomal dipeptidase, which was GPI membrane-anchored. Protein-kinase activities, mainly serine-kinase, were enriched during the DRM purification. Using the typical FTIR olefinic =C–H bands of the acyl chains, a global decrease in the unsaturation level of DRM lipids was observed as compared with total membranes. Three main phospholipids were identified in DRM. Their fatty acid compositions were determined by gas chromatography and compared with those of total membranes. The most enriched saturated fatty acid was palmitic acid (+44% for phosphatidylethanolamine, +52% for phosphatidylcholine and +49% for sphingomyelin), agreeing with a selection of specific phospholipids among the saturated ones during the DRM purification. |
| Databáze: | OpenAIRE |
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