Chaperone-assisted folding of a single-chain antibody in a reconstituted translation system |
| |
Authors: | Ying Bei-Wen Taguchi Hideki Ueda Hiroshi Ueda Takuya |
| |
Institution: | Department of Medical Genome Sciences, Graduate School of Frontier Sciences, Kashiwa, Chiba, Japan. |
| |
Abstract: | A protein-synthesizing system based on a minimal set of purified components was used to investigate the roles molecular chaperones play in the folding of newly synthesized polypeptides. After we ascertained that this system lacks intrinsic chaperones, the effect of adding chaperones in a co-translational or post-translational manner was directly evaluated. An aggregation-prone single-chain antibody was used as the model nascent chain. The participation of the trigger factor or the DnaK system during translation efficiently increased the level of functional protein that was generated. In addition, both systems also acted as chaperones after translation had been stopped. In contrast, the GroEL/ES system showed little or no co- or post-translational assistance in folding. |
| |
Keywords: | Molecular chaperone Protein folding Cell-free translation Co-translational Post-translational scFv |
本文献已被 ScienceDirect PubMed 等数据库收录! |