| PPC106 | |
| ID |  PPC106 |
| Project ID | PPC1 [Protein] [PREIMS] |
| Subject | Synthesis of membrane protein that uses liposome coexistent wheat cell-free protein synthesis method |
| Project Theme | Development of Advanced Production Technologies for Target Proteins |
| Principal Investigator | Shigeyuki Yokoyama |
| Affiliation | Systems and Structural Biology Center, Yokohama Institute, RIKEN |
| Backgrounds | - The extract from extensively washed wheat embryo particles can provide a very productive cell-free protein synthesis system A useful method taking advantage of the wheat cell-free system has been sought for preparation of variety type of membrane proteins |
| Highlights | - We synthesized a functional AtPPT1 plant phosphate transporter using a wheat cell-free system in the presence of liposomes Furthermore, using a liposome-supplemented wheat cell-free translation system, 40 mammalian membrane proteins were synthesized in their functionally active forms with high efficiency |
| Outline | Cell-free protein synthesis systems using a variety of cell extracts have emerged as a promising tool for protein production. Three types of methods have been employed in the cell-free synthesis of functional membrane proteins; 1) refolding and solubilization of translation products, 2) translation in the presence of detergents, 3) translation in the presence of liposomes. We applied a liposome-supplemented wheat cell-free translation system to the production of membrane proteins. Lipid bilayer liposomes were prepared from asolectin, purified phospholipid. First, we synthesized a functional AtPPT1 plant phosphate transporter and confirmed its high transport activity. To test whether co-translational association with liposomes is observed in the synthesis of other membrane proteins, we tried comprehensive synthesis of mammalian membrane proteins. Using this method, 40 mammalian membrane proteins were efficiently associated with liposomes co-translationally. Therefore, this synthesis method is useful in the preparation of membrane protein for structural and functional analysis. |
| Application | FBB5 MPA2 MPA6 |
| Article | - |
| CSML File | PPC106.csml |