| Nov 24, 2023 |
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(Nanowerk News) A recent study published in Nature Methods (“Peptide sequencing based on host–guest interaction-assisted nanopore sensing”) showcases the development of a revolutionary protein sequencing technology, promising significant advances in the fields of proteomics and clinical diagnostics.
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This research, led by Prof. Haichen Wu from the Institute of Chemistry, Chinese Academy of Sciences (CAS), and Prof. Lei Liu from the Institute of High Energy Physics, CAS, alongside their collaborators, introduces a novel peptide sequencing method employing host–guest interaction-assisted nanopore sensing.
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Protein sequencing has evolved since the 1950s, with the landmark determination of insulin’s amino acid sequence by Sanger. Traditionally, mass spectrometry and Edman degradation have been the primary methods for protein sequencing.
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Nanopore sensing, emerging as a groundbreaking single-molecule technique in recent decades, has been instrumental in advancing DNA sequencing. This success has motivated efforts to adapt nanopore technology for protein sequencing, a challenging endeavor due to the need for unidirectional transport of heterogeneously charged peptide chains through a nanopore and the electrical identification of the 20 amino acids.
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In their innovative approach, the research team uses an enhanced host-guest interaction-assisted nanopore sensing strategy. The process involves initially digesting model peptides with carboxypeptidases to produce a mixture of amino acids. These amino acids are then covalently linked to a FGGCD8⊂CB[7] peptide probe and passed through wildtype α-hemolysin or its mutants in translocation experiments. The current blockage caused by each FGGC(X)D8⊂CB[7] peptide helps identify the amino acid X, and their relative abundance helps determine the peptide sequence.
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This breakthrough study demonstrates a promising new method for accurately determining peptide amino acid sequences. Despite certain limitations, it represents a significant step forward in protein sequencing and opens new possibilities for future advancements in the field.
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