| Autor: |
Drachmann, Nikolaj Düring, Olesen, Claus |
| Jazyk: |
angličtina |
| Rok vydání: |
2016 |
| Předmět: |
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| Zdroj: |
Drachmann, N D & Olesen, C 2016, Lipid exchange by ultracentrifugation . in P-Type ATPases. Methods and Protocols . Springer, New York, Methods in Molecular Biology, vol. 1377, pp. 397-402 . |
| Popis: |
Lipids play an important role in maintaining P-type ATPase structure and function, and often they are crucial for ATPase activity. When the P-type ATPases are in the membrane, they are surrounded by a mix of different lipids species with varying aliphatic chain lengths and saturation, and the complex interplay between the lipids and the P-type ATPases are still not well understood. We here describe a robust method to exchange the majority of the lipids surrounding the ATPase after solubilisation and/or purification with a target lipid of interest. The method is based on an ultracentrifugation step, where the protein sample is spun through a dense buffer containing large excess of the target lipid, which results in an approximately 80-85 % lipid exchange. The method is a very gently technique that maintains protein folding during the process, hence allowing further characterization of the protein in the presence of a target lipid of interest. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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