Tuning the conformation of G-quadruplexes by sodium and potassium ions: application to photometric and fluorometric determination of amyloid β(1–40)

Abstract

A dual channel method is described for the determination of the amyloid-β peptide Aβ(1–40) that is associated with Alzheimer’s disease. The method exploits (a) conformational changes of a G-quadruplex that are triggered by Na⁺ and K⁺ ions and (b) the strong affinity between Aβ(1–40) and Cu²⁺. A G-quadruplex DNA forms an antiparallel structure in the presence of Na⁺ and can catalyze the oxidation of tetramethylbenzidine by H₂O₂ system in the presence of Cu²⁺ to form a visible blue color. If, however, Cu²⁺ binds to Aβ(1–40), the blue color is no longer formed. Measuring the absorption decrease at 452 nm, the determination of Aβ(1–40) is realized. If K⁺ is added to the Na⁺-containing buffer, the antiparallel G-quadruplex DNA is transformed to parallel. This leads to the insertion of protoporphyrin IX (PPIX) into the G-quadruplex and generates enhanced fluorescent signal, with excitation/emission wavelength at 410/630 nm. The G-quadruplex then catalyzes the metalation of PPIX by Cu²⁺, and the fluorescence intensity decreases. In the presence of Aβ(1–40), the formation of Aβ(1–40)-Cu²⁺ triggers the recovery of the fluorescence. The Na⁺/K⁺-induced tuning of the conformation of the G-quadruplex with the same sequence enables dual (colorimetric and fluorometric) determination of Aβ(1–40), with detection limits of 4.9 pM and 2.3 pM, respectively. The cost is quite low since the developed strategy is label free and enzyme free by using low-cost DNA and Cu²⁺. More importantly, the dual channel determination operation is very simple without any further modification process. Graphical abstract Tuning the conformation of G-quadruplexes by sodium(I) and potassium(I): application to photometric and fluorometric determination of amyloid β(1–40).