Multi-pass, single-molecule nanopore reading of long protein strands

The ability to sequence single protein molecules in their native, full-length form would enable a more comprehensive understanding of proteomic diversity. Current technologies, however, are limited in achieving this goal1,2. Here, we establish a method for the long-range, single-molecule reading of intact protein strands on a commercial nanopore sensor array. By using the ClpX unfoldase to ratchet proteins through a CsgG nanopore3,4, we provide single-molecule evidence that ClpX translocates substrates in two-residue steps. This mechanism achieves sensitivity to single amino acids on synthetic protein strands hundreds of amino acids in length, enabling the sequencing of combinations of single-amino-acid…

• NS
  National Science Foundation

  Awards: EF-2021552, 2021552
• JA
  Japan Agency for Medical Research and Development
• ON
  Oxford Nanopore Technologies
• NI
  National Institutes of Health
• JS
  Japan Society for the Promotion of Science