# Label-free optical detection of single enzyme-reactant reactions and associated conformational changes

## TL;DR

This paper introduces a novel label-free optical method using plasmonic nanorods and microcavities to observe enzyme-reactant interactions and conformational changes at the single-molecule level, avoiding the need for molecular labels.

## Contribution

The study demonstrates a new optical technique combining plasmonic nanorods and whispering gallery mode microcavities for real-time, label-free detection of enzyme activity and structural dynamics at the single-molecule scale.

## Key findings

- Able to distinguish low and high polymerase activities via signal characteristics
- Detected conformational changes in polymerase molecules
- Provided thermodynamic insights into DNA polymerization processes

## Abstract

Monitoring the kinetics and conformational dynamics of single enzymes is crucial in order to better understand their biological functions as these motions and structural dynamics are usually unsynchronized among the molecules. Detecting the enzyme-reactant interactions and associated conformational changes of the enzyme on a single molecule basis, however, remain as a challenge with established optical techniques due to the commonly required labeling of the reactants or the enzyme itself. The labeling process is usually non-trivial and the labels themselves might skew the physical properties of the enzyme. Here we demonstrate an optical, label-free method capable of observing enzymatic interactions and the associated conformational changes on the single molecule level. We monitor polymerase/DNA interactions via the strong near-field enhancement provided by plasmonic nanorods resonantly coupled to whispering gallery modes in microcavities. Specifically, we employ two different recognition schemes: one in which the kinetics of polymerase/DNA interactions are probed in the vicinity of DNA-functionalized nanorods, and the other in which these interactions are probed via the magnitude of conformational changes in the polymerase molecules immobilized on nanorods. In both approaches we find that low and high polymerase activities can be clearly discerned via their characteristic signal amplitude and signal length distributions. Furthermore, the thermodynamic study of the monitored interactions suggests the occurrence of DNA polymerization. This work constitutes a proof-of-concept study of enzymatic activities via plasmonically enhanced microcavities and establishes an alternative and label-free method capable of investigating structural changes in single molecules.