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Single-molecule sensing inside stereo- and regio-defined hetero-nanopores.
Liu, Wei; Zhu, Qiang; Yang, Chao-Nan; Fu, Ying-Huan; Zhang, Ji-Chang; Li, Meng-Yin; Yang, Zhong-Lin; Xin, Kai-Li; Ma, Jing; Winterhalter, Mathias; Ying, Yi-Lun; Long, Yi-Tao.
Afiliación
  • Liu W; School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, China.
  • Zhu Q; Molecular Sensing and Imaging Center, Nanjing University, Nanjing, China.
  • Yang CN; School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, China.
  • Fu YH; School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, China.
  • Zhang JC; Molecular Sensing and Imaging Center, Nanjing University, Nanjing, China.
  • Li MY; School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, China.
  • Yang ZL; Molecular Sensing and Imaging Center, Nanjing University, Nanjing, China.
  • Xin KL; School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, China.
  • Ma J; Molecular Sensing and Imaging Center, Nanjing University, Nanjing, China.
  • Winterhalter M; School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, China.
  • Ying YL; Molecular Sensing and Imaging Center, Nanjing University, Nanjing, China.
  • Long YT; Chemistry and Biomedicine Innovation Center, Nanjing University, Nanjing, China.
Nat Nanotechnol ; 2024 Aug 20.
Article en En | MEDLINE | ID: mdl-39164412
ABSTRACT
Heteromeric pore-forming proteins often contain recognition patterns or stereospecific selection filters. However, the construction of heteromeric pore-forming proteins for single-molecule sensing is challenging due to the uncontrollability of producing position isomers and difficulties in purification of regio-defined products. To overcome these preparation obstacles, we present an in situ strategy involving single-molecule chemical modification of a heptameric pore-forming protein to build a stereo- and regio-specific heteromeric nanopore (hetero-nanopore) with a subunit stoichiometric ratio of 34. The steric hindrance inherent in the homo-nanopore of K238C aerolysin directs the stereo- and regio-selective modification of maleimide derivatives. Our method utilizes real-time ionic current recording to facilitate controlled voltage manipulation for stoichiometric modification and position-based side-isomer removal. Single-molecule experiments and all-atom molecular dynamics simulations revealed that the hetero-nanopore features an asymmetric stereo- and regio-defined residue structure. The hetero-nanopore produced was characterized by mass spectrometry and single-particle cryogenic electron microscopy. In a proof-of-concept single-molecule sensing experiment, the hetero-nanopore exhibited 95% accuracy for label-free discrimination of four peptide stereoisomers with single-amino-acid structural and chiral differences in the mixtures. The customized hetero-nanopores could advance single-molecule sensing.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Nat Nanotechnol Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Nat Nanotechnol Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido