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Real-time label-free direct electronic monitoring of topoisomerase enzyme binding kinetics on graphene.

MPG-Autoren
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Yu,  H. K.
Department of Dynamics at Surfaces, MPI for Biophysical Chemistry, Max Planck Society;

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Zitation

Zuccaro, L., Tesauro, C., Kurkina, T., Fiorani, P., Yu, H. K., Knudsen, B. R., et al. (2015). Real-time label-free direct electronic monitoring of topoisomerase enzyme binding kinetics on graphene. ACS Nano, 9(11), 11166-11176. doi:10.1021/acsnano.5b05709.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-0029-3312-D
Zusammenfassung
Monolayer graphene field-effect sensors operating in liquid have been widely deployed for detecting a range of analyte species often under equilibrium conditions. Here we report on the real-time detection of the binding kinetics of the essential human enzyme, topoisomerase I interacting with substrate molecules (DNA probes) that are immobilized electrochemically on to monolayer graphene strips. By monitoring the field-effect characteristics of the graphene biosensor in real-time during the enzyme–substrate interactions, we are able to decipher the surface binding constant for the cleavage reaction step of topoisomerase I activity in a label-free manner. Moreover, an appropriate design of the capture probes allows us to distinctly follow the cleavage step of topoisomerase I functioning in real-time down to picomolar concentrations. The presented results are promising for future rapid screening of drugs that are being evaluated for regulating enzyme activity.