Protein and lipid fingerprinting of native-like membrane complexes by combining thin layer chromatography and protein electrophoresis: the example of lung surfactant
Traditionally, thin layer chromatography (TLC) has been used for the analysis of lipids isolated from membrane complexes. Here, we describe a method based on the combination of TLC and SDS-polyacrylamide gel electrophoresis (SDS-PAGE) for the qualitative analysis of the protein/lipid profile of memb...
| Autores: | , , , |
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| Tipo de recurso: | artículo |
| Fecha de publicación: | 2019 |
| País: | España |
| Institución: | Universidad Complutense de Madrid (UCM) |
| Repositorio: | Docta Complutense |
| Idioma: | inglés |
| OAI Identifier: | oai:docta.ucm.es:20.500.14352/13523 |
| Acceso en línea: | https://hdl.handle.net/20.500.14352/13523 |
| Access Level: | acceso abierto |
| Palabra clave: | 577.1 577.2 Diagnostic tools Lipoproteins Phospholipids Proteomics Pulmonary surfactant Biología molecular (Biología) Bioquímica (Biología) 2415 Biología Molecular 2302 Bioquímica |
| Sumario: | Traditionally, thin layer chromatography (TLC) has been used for the analysis of lipids isolated from membrane complexes. Here, we describe a method based on the combination of TLC and SDS-polyacrylamide gel electrophoresis (SDS-PAGE) for the qualitative analysis of the protein/lipid profile of membrane complexes such as those of lung surfactant. For this purpose, native lung surfactant was applied onto a silica TLC plate in the form of an aqueous suspension, preserving not only hydrophilic proteins associated to lipids, but also native protein-lipid interactions. Using native membrane complexes in a TLC allows the differential migration of lipids and their separation from the protein components. As a result, (partly) delipidated protein-enriched bands can be visualized and analysed by SDS-PAGE to identify proteins originally associated with lipids. Interestingly, the hydrophobic surfactant protein SP-C, which interact tightly with lipids in native membrane complexes, migrate through the TLC plate configuring specific bands that differ from those corresponding to lipids or proteins. This method therefore allows the detection and analysis of strong native-like protein-lipid interactions. |
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