Mechanism of a Disassembly Driven Sensing System Studied by Stopped-Flow Kinetics

15 April 2021, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Mechanistic studies were carried out on the kinetics for the assembly of a DimerDye (DD12) and the binding of the monomeric DimerDye (DD1) with nicotine in aqueous buffer and artificial saliva. DD12 is non-fluorescent, while monomeric DD1 and DD1-nicotine fluoresce. Binding isotherms were determined from steady-state fluorescence experiments. The report includes measurements of the steady-state fluorescence at pHs 2.2, 6.3 and 12.1, and stopped-flow kinetic data for the homodimerization forming DD12 and DD1-nicotine formation in buffer and artificial saliva. Analysis of the homodimerization kinetics led to the recovery of the association and dissociation rate constants for DD12. These rate constants were used in the global analysis for the coupled kinetics for DD1-nicotine formation, which led to the determination of the association and dissociation rate constants for nicotine binding to DD1.

Keywords

Disassembly sensing
DimerDye
Supramolecular dynamics
Kinetics
Calixarene
Flurorescence
Saliva
Self-assembly

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