A Toolbox of Chromones and Quinolones for Measuring a Wide Range of ATP Concentrations

Abstract

A series of 26 3-hydroxychromones, three bis-flavonols and four 3-hydroxyquinolones were studied to evaluate their fluorescence response to interaction with ATP in buffer. The dyes differ by the total charge, the size and number of their aromatic units, as well as the position or electron-donating ability of their substituents. All of them were suggested to form complexes with ATP of 1:1 and 1:2 stoichiometry, which can be evidenced by their bright fluorescence and their 3000-6000 cm red-shifted excitation band. These fluorescent complexes allow detection of ATP concentrations over 3 orders of magnitude, whereas most other known probes cover no more than two orders. In total, the dyes allow ATP detection from 0.001 to 57 mm. In addition, most of the dye-ATP complexes can be excited in the visible and monitored in the red region of the spectrum. The response amplitude of the described dyes to ATP is as high as for the best known probes. Considering that complexation takes place at neutral pH, the studied dyes form a toolbox of fluorescent probes for intensiometric and ratiometric measurements of ATP concentration in a broad concentration range. Finally, the obtained results stimulate the idea that most of natural 3-hydroxyflavones in living cells may form complexes with ATP.

Molecular Receptors for Recognition and Sensing of Nucleotides

“…[18] Pivovarenko and co-workersd iscovered that 3-hydroxyflavones could bind and detect ATPi na queous solution in ab road concentrationr ange. [19] Among al argen umber of the investigated chromones and quinolones,t he authors found that hydroxyland dialkylamino-derivatives (18,19)d isplay an ew bathochromically shifted band in the presence of ATPi na queous solution. Ther ecognition process is accompanied by as trong fluorescencei ncrease (30-50-fold).I ti sp roposed that adenine interacts with the dye, releasing the water from the dye, whereas the OH groups can form hydrogen bonds with basic atoms of the nucleotide.…”

“…Interestingly, amphiphilic dye 17 can also form self‐assembled structures but with a micelle‐like structure, which can bind ATP on its outer surface resulting in fluorescence enhancement . Pivovarenko and co‐workers discovered that 3‐hydroxyflavones could bind and detect ATP in aqueous solution in a broad concentration range . Among a large number of the investigated chromones and quinolones, the authors found that hydroxyl‐ and dialkylamino‐derivatives ( 18 , 19 ) display a new bathochromically shifted band in the presence of ATP in aqueous solution.…”

An uracil-linked hydroxyflavone probe for the recognition of ATP

“…The interaction of DMHF with nucleotides and a computer aided simulation on the geometry of DMHF∙ATP complexes in vacuo was also reported. A recent study by the same authors describe further flavones with various association constants [ 45 ], however, none of them are the result of rational design.…”

Design and Synthesis of Molecular Tweezer Based on 3-Hydroxyflavone for the Detection of Atp

“…Ратіометричні зонди містять у своєму складі кілька додаткових елементів, таких як донор та акцептор енергії (у FRET-зондах [22,24]) або два однакові флуорофори, як у ексимерних зондах [22]. Важливим кроком на шляху до спрощення структури АТФ-зондів є розробка конструкцій на основі флавонолу та його ізостер: 3-гідрокси-4'-(диметиламіно)флавон (FME) [28,29] і похідні хромону та хінолону [30]. Зонд FME, синтетичний аналог природних флавонолів [31,32], незважаючи на просту будову, дає високоселективну флуоресцентну відповідь при зв'язуванні з АТФ порівняно з іншими нуклеотидами у водних буферних розчинах [29].…”

“…З допомогою цього зонду була здійснена реєстрація виділення АТФ мітохондріями під впливом сукцинату in vitro [28]. Остання публікація [30] свідчить, що 31 з 33 досліджених флавонів та хінолонів різної будови та заряду дають потужну флуоресцентну відповідь при зв'язуванні з АТФ і можуть бути ефективно застосовані у водних розчинах для виявлення АТФ у концентраційних межах 1-50000 мкМ.…”