Single Molecule Thermodynamic Penalties Applied to Enzymes by Whispering Gallery Mode Biosensors

Optical microcavities, particularly whispering gallery mode (WGM) microcavities enhanced by plasmonic nanorods, are emerging as powerful platforms for single-molecule sensing. However, the impact of optical forces from the plasmonic near field on analyte molecules is inadequately understood. Using a standard optoplasmonic WGM single-molecule sensor to monitor two enzymes, both of which undergo an open-to-closed-to-open conformational transition, the work done on an enzyme by the WGM sensor as atoms of the enzyme move through the electric field gradient of the plasmonic hotspot during conformational change has been quantified. As the work done by the sensor on analyte enzymes can be modulated by varying WGM intensity, the WGM microcavity system can be used to apply free energy penalties to regulate enzyme activity at the single-molecule level. The findings advance the understanding of optical forces in WGM single-molecule sensing, potentially leading to the capability to precisely manipulate enzyme activity at the single-molecule level through tailored optical modulation. Turnover of enzymes bound to optoplasmonic sensors can be detected at the single-molecule level. By modulating the strength of the electric field generated by the plasmonic gold nanoparticle, optoplasmonic sensors may be capable of generating forces that oppose turnover and that could therefore be used to control enzymes at the single molecule level. Created with BioRender.com image