PMID:16138309
| Citation |
Lorch, M, Faham, S, Kaiser, C, Weber, I, Mason, AJ, Bowie, JU and Glaubitz, C (2005) How to prepare membrane proteins for solid-state NMR: A case study on the alpha-helical integral membrane protein diacylglycerol kinase from E. coli. Chembiochem 6:1693-700 |
|---|---|
| Abstract |
Several studies have demonstrated that it is viable to use microcrystalline preparations of water-soluble proteins as samples in solid-state NMR experiments [1-5]. Here, we investigate whether this approach holds any potential for studying water-insoluble systems, namely membrane proteins. For this case study, we have prepared proteoliposomes and small crystals of the alpha-helical membrane-protein diacylglycerol kinase (DGK). Preparations were characterised by 13C- and 15N-cross-polarization magic-angle spinning (CPMAS) NMR. It was found that crystalline samples produce better-resolved spectra than proteoliposomes. This makes them more suitable for structural NMR experiments. However, reconstitution is the method of choice for biophysical studies by solid-state NMR. In addition, we discuss the identification of lipids bound to membrane-protein crystals by 31P-MAS NMR. |
| Links |
PubMed Online version:10.1002/cbic.200500054 |
| Keywords |
Adenosine Triphosphate/pharmacology; Crystallization; Diacylglycerol Kinase/chemistry; Diacylglycerol Kinase/metabolism; Escherichia coli/enzymology; Lipids/chemistry; Membrane Proteins/chemistry; Membrane Proteins/metabolism; Nuclear Magnetic Resonance, Biomolecular/methods; Protein Structure, Secondary |
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