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Title
Japanese:Regulated N-Terminal Modification of Proteins Synthesized Using a Reconstituted Cell-Free Protein Synthesis System. 
English:Regulated N-Terminal Modification of Proteins Synthesized Using a Reconstituted Cell-Free Protein Synthesis System. 
Author
Japanese: Matsumoto R, 丹羽達也, Shimane Y, Kuruma Y, Hideki Taguchi, Takashi Kanamori.  
English: Matsumoto R, Niwa T, Shimane Y, Kuruma Y, Hideki Taguchi, Takashi Kanamori.  
Language English 
Journal/Book name
Japanese:ACS synthetic biology 
English:ACS synthetic biology 
Volume, Number, Page        
Published date June 16, 2023 
Publisher
Japanese: 
English: 
Conference name
Japanese: 
English: 
Conference site
Japanese: 
English: 
Official URL https://doi.org/10.1021/acssynbio.3c00191
 
DOI https://doi.org/10.1021/acssynbio.3c00191
Abstract The N-terminal modification of nascent proteins, such as acetylation and myristoylation, is one of the most abundant post-translational modifications. To analyze the function of the modification, it is important to compare the modified and unmodified proteins under defined conditions. However, it is technically difficult to prepare unmodified proteins because cell-based systems contain endogenous modification systems. In this study, we developed a cell-free method to conduct N-terminal acetylation and myristoylation of nascent proteins in vitro using a reconstituted cell-free protein synthesis system (PURE system). Proteins synthesized using the PURE system were successfully acetylated or myristoylated in a single-cell-free mixture in the presence of modifying enzymes. Furthermore, we performed protein myristoylation in giant vesicles, which resulted in their partial localization to the membrane. Our PURE-system-based strategy is useful for the controlled synthesis of post-translationally modified proteins.

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