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Issue 3, 2019
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Fluorescence correlation spectroscopy for multiple-site equilibrium binding: a case of doxorubicin–DNA interaction

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Abstract

Quantitative description of the interaction between doxorubicin (DOX), a broadly used anticancer drug, and DNA is the key to understand the action mechanism and side effects of its clinical use. However, the reported equilibrium constants of DOX–DNA interaction obtained using a range of different analytical methods vary even by several orders of magnitude. Herein, we propose a novel application of a single-molecule technique – fluorescence correlation spectroscopy (FCS) – to probe the interaction between DOX and two types of DNA (pUC19 and calf thymus DNA), taking advantage of intrinsic self-fluorescence of DOX. We provide an analytical formula for autocorrelation analysis to determine the equilibrium constant of DOX–DNA complex-formation, where binding of multiple DOX molecules to a DNA chain is included in the reaction–diffusion model. Our FCS-based method not only quantitatively revealed the values of equilibrium constant, but also implied that the stability of DOX–DNA complex is related to the types of base pair rather than the length or structure of the DNA. This work opens a promising pathway toward quantitative determination of molecular interactions in complex systems such as living cells or organisms at single-molecule level.

Graphical abstract: Fluorescence correlation spectroscopy for multiple-site equilibrium binding: a case of doxorubicin–DNA interaction

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Supplementary files

Article information


Submitted
30 Oct 2018
Accepted
18 Dec 2018
First published
08 Jan 2019

This article is Open Access

Phys. Chem. Chem. Phys., 2019,21, 1572-1577
Article type
Paper

Fluorescence correlation spectroscopy for multiple-site equilibrium binding: a case of doxorubicin–DNA interaction

X. Zhang, A. Poniewierski, K. Sozański, Y. Zhou, A. Brzozowska-Elliott and R. Holyst, Phys. Chem. Chem. Phys., 2019, 21, 1572
DOI: 10.1039/C8CP06752J

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