Size-tunable transmembrane nanopores assembled from decomposable molecular templates

Area of Science:

• Biochemistry
• Biophysics
• Materials Science

Background:

• Transmembrane nanopores are crucial for molecular transport and sensing.
• Their precise size, particularly inner diameter, dictates sensing capabilities and applications.
• Natural nanopore assembly involves multiple monomers within lipid bilayers.

Purpose of the Study:

• To introduce a novel template molecule-aided assembly strategy for constructing size-tunable transmembrane nanopores.
• To demonstrate the ability to control nanopore size and function artificially.
• To enable applications in single-molecule sensing and analysis.

Main Methods:

• Utilized cyclodextrins of varying sizes as templates to guide the assembly of the α-helical barreled peptide (Wza).
• Assembled functional nanopores capable of insertion into lipid bilayers, forming stable single channels.
• Employed enzyme digestion to remove cyclodextrin templates, preserving nanopore integrity and function.
• Synthesized peptides and performed chemical modifications for functional modulation.

Main Results:

• Successfully constructed size-tunable transmembrane protein nanopores ranging from 1.1 to 1.8 nm in diameter.
• Achieved stable insertion of assembled nanopores into lipid bilayers for single-molecule sensing.
• Demonstrated that removed templates do not compromise nanopore structure or function.
• Showcased facile modulation of pore function through chemical synthesis and modification.

Conclusions:

• The template molecule-aided assembly strategy offers a simple and convenient method for artificial synthesis of transmembrane protein nanopores.
• Nanopore size is precisely controllable by selecting the appropriate template molecule.
• This approach allows for easy modulation of pore function without genetic manipulation, broadening potential applications in biosensing.