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Real-time measurement of protein-protein interactions at single-molecule resolution using a biological nanopore.

Avinash Kumar ThakurLiviu Movileanu
Published in: Nature biotechnology (2018)
Protein-protein interactions (PPIs) are essential for many cellular processes. However, transient PPIs are difficult to measure at high throughput or in complex biological fluids using existing methods. We engineered a genetically encoded sensor for real-time sampling of transient PPIs at single-molecule resolution. Our sensor comprises a truncated outer membrane protein pore, a flexible tether, a protein receptor and a peptide adaptor. When a protein ligand present in solution binds to the receptor, reversible capture and release events of the receptor can be measured as current transitions between two open substates of the pore. Notably, the binding and release of the receptor by a protein ligand can be unambiguously discriminated in a complex sample containing fetal bovine serum. Our selective nanopore sensor could be applied for single-molecule protein detection, could form the basis for a nanoproteomics platform or might be adapted to build tools for protein profiling and biomarker discovery.
Keyphrases
  • single molecule
  • binding protein
  • high throughput
  • living cells
  • atomic force microscopy
  • protein protein
  • amino acid
  • small molecule
  • single cell
  • minimally invasive
  • subarachnoid hemorrhage