2025-06-20 海洋研究開発機構,ジーンフロンティア株式会社,東京科学大学
図1 本研究で開発した無細胞系の概要図. 再構築型の無細胞系PURE systemで合成したVHH抗体はTEVプロテアーゼでトリミングを受けた後、 ミリストイル基転移酵素によりミリストイルCoA(脂質源)またはパルミトイルCoAから脂質を修飾される。 これにより疎水的相互作用が働きリポソーム表面に固定される。抗体を固定したリポソームは標的となるがん細胞に結合する。
<関連情報>
- https://www.jamstec.go.jp/j/about/press_release/20250620/
- https://pubs.acs.org/doi/10.1021/acssynbio.5c00155
無細胞系における脂質の修飾とタンパク質の膜局在化 Lipid Modification and Membrane Localization of Proteins in Cell-Free System
Rena Matsumoto,Tatsuya Niwa,Kaori Kuno,Yasuhiro Shimane,Yutetsu Kuruma,and Takashi Kanamori
ACS Synthetic Biology Published: June 19, 2025
DOI:https://doi.org/10.1021/acssynbio.5c00155
Abstract
Post-translational modifications are an essential process for proper protein function and localization. In particular, lipid modification plays a crucial role in the spatial regulation of proteins functioning on a lipid membrane surface. While cell-free protein synthesis allows rapid protein production, technical advances in lipidation modification are behind. Here, we developed a cell-free system for the myristoylation and palmitoylation of proteins. Based on our previous study, we improved myristoylation efficiency by trimming a precursor nascent peptide, which undergoes lipidation at the N-terminal glycine. We also found that N-myristoyltransferase (NMT) catalyzes both myristoylation and palmitoylation. The localization of lipidated proteins onto liposomes is further aided by the insertion of polyarginine residues downstream of the NMT recognition site. Finally, we demonstrated that lipidation of VHH antibodies and localization onto liposomes resulted in target-specific binding to cancer cells. This system offers a platform for displaying soluble proteins on lipid membranes, with potential applications in developing liposomes for targeted cell binding.
