PMID:20194040
| Citation |
Abdine, A, Verhoeven, MA, Park, KH, Ghazi, A, Guittet, E, Berrier, C, Van Heijenoort, C and Warschawski, DE (2010) Structural study of the membrane protein MscL using cell-free expression and solid-state NMR. J. Magn. Reson. 204:155-9 |
|---|---|
| Abstract |
High-resolution structures of membrane proteins have so far been obtained mostly by X-ray crystallography, on samples where the protein is surrounded by detergent. Recent developments of solid-state NMR have opened the way to a new approach for the study of integral membrane proteins inside a membrane. At the same time, the extension of cell-free expression to the production of membrane proteins allows for the production of proteins tailor made for NMR. We present here an in situ solid-state NMR study of a membrane protein selectively labeled through the use of cell-free expression. The sample consists of MscL (mechano-sensitive channel of large conductance), a 75kDa pentameric alpha-helical ion channel from Escherichia coli, reconstituted in a hydrated lipid bilayer. Compared to a uniformly labeled protein sample, the spectral crowding is greatly reduced in the cell-free expressed protein sample. This approach may be a decisive step required for spectral assignment and structure determination of membrane proteins by solid-state NMR. |
| Links |
PubMed Online version:10.1016/j.jmr.2010.02.003 |
| Keywords |
Cell-Free System/chemistry; Computer Simulation; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/ultrastructure; Ion Channels/chemistry; Ion Channels/ultrastructure; Magnetic Resonance Spectroscopy/methods; Membrane Proteins/chemistry; Membrane Proteins/ultrastructure; Models, Chemical; Models, Molecular; Powders |
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