合成が難しいナノポア形成ベータシートペプチドを 無細胞たんぱく質合成系によって簡便に合成することに成功 ～1分子をラベルフリーに検出可能なバイオセンシング技術への応用に期待～

2026-09-16 東京農工大学,科学技術振興機構

図1：本研究の概要。ナノポア形成ベータシートペプチドを、リポソームを添加した再構成型無細胞合成系を用いて合成した（左）。脂質組成の最適化によりナノポア形成に成功し、この無細胞合成により合成したSVG28ナノポアにポリペプチドを添加すると、通過するポリペプチドに由来する電流値の減少が観察された（右）。

＜関連情報＞

• https://www.jst.go.jp/pr/announce/20260916/index.html
• https://www.jst.go.jp/pr/announce/20260916/pdf/20260916.pdf
• https://pubs.acs.org/ancac3/article/doi/10.1021/acsnano.5c15163/5430674/Liposome-Supplemented-Cell-free-Synthesis-of-a

リポソームを添加した無細胞系によるβバレル型ナノポアを形成するペプチドの合成 Liposome-Supplemented Cell-free Synthesis of a Peptide that Forms β-barrel Nanopores

Shoko Fujita;Mana Sato;Ryuji Kawano
ACS Nano  Published:September 15, 2026
DOI:https://doi.org/10.1021/acsnano.5c15163

Abstract

Rational design of nanopore-forming proteins and peptides is a promising approach for single-molecule analysis by using custom-designed nanopores tailored to specific target molecules. One challenge in this approach is the expression or synthesis of the designed nanopores, due to their high hydrophobicity. We sought to streamline the synthesis of an artificial and highly aggregative nanopore-forming peptide using liposome-supplemented cell-free protein synthesis. By optimizing the liposomal lipid composition, we recovered yields of the expressed aggregative peptide that were suitable for functional measurements. Channel current measurements involving peptide-embedded liposome fusion to a planar lipid bilayer confirmed the pore formation and single-molecule detection capabilities of the expressed peptide nanopore. Through the exploration of various lipid compositions, it was found that the incorporation of anionic lipids and a sufficient acyl chain length were crucial for efficient peptide insertion and the reproduction of native oligomeric states. Such modulation of liposomal lipid composition alongside cell-free protein synthesis has the capacity to alleviate the aggregation propensity constraints in the design of nanopores and other membranous peptides.