Single-Molecule Protein Interactions and Unfolding Revealed by Plasmon-Enhanced Fluorescence.

ABSTRACT: Single-molecule characterization of protein interaction kinetics can unravel crucial mechanisms that are averaged out with ensemble-average approaches. However, current approaches based on single-molecule fluorescence are limited in terms of signal brightness and time resolution. We introduce a novel platform to quantify protein-protein interactions at the single-molecule level using plasmon-enhanced fluorescence microscopy. We illustrate the power of this approach using PDZ protein that is conjugated to plasmonic particles using a novel DNA-mediated hybridization method that provides spatial and orientational control over the proteins' immobilization. Single-molecule kinetic studies uncover heterogeneities in the interaction where a subpopulation of events exhibits a distinct bound-state-

SUBMITTER: Teeuwen RWH 

PROVIDER: S-EPMC12311895 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature