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

Avinash Kumar Thakur, Liviu Movileanu

Research output: Contribution to journalLetter

8 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)96-104
Number of pages9
JournalNature Biotechnology
Volume37
Issue number1
DOIs
StatePublished - Jan 1 2019

Fingerprint

Nanopores
Time measurement
Proteins
Molecules
Sensors
Ligands
Peptide Receptors
Biomarkers
Membrane Proteins
Peptides
Throughput
Sampling
Fluids
Membranes
Serum

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Cite this

Real-time measurement of protein–protein interactions at single-molecule resolution using a biological nanopore. / Thakur, Avinash Kumar; Movileanu, Liviu.

In: Nature Biotechnology, Vol. 37, No. 1, 01.01.2019, p. 96-104.

Research output: Contribution to journalLetter

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