کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5406712 | 1393185 | 2010 | 5 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: CommunicationStructural study of the membrane protein MscL using cell-free expression and solid-state NMR CommunicationStructural study of the membrane protein MscL using cell-free expression and solid-state NMR](/preview/png/5406712.png)
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 75 kDa pentameric α-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.
Journal: Journal of Magnetic Resonance - Volume 204, Issue 1, May 2010, Pages 155-159