A switch-on luminescent europium(iii) probe for selective and time-resolved detection of adenosine diphosphate (ADP)

Abstract

Adenosine diphosphate (ADP) is a key product of two essential classes of biological reactions, catalysed by ATPases and kinases. This makes ADP a highly appealing target for supramolecular detection. However, doing so selectively is exceedingly difficult due to ADP's lower overall charge and similar structure to ATP and the need for compatibility with biological media. Overcoming this challenge, here we present a water-soluble, ADP-selective, luminescent europium(III) probe suitable for use in vitro and in cellular microscopy. This negatively charged Eu(III) complex binds ADP reversibly and responds by switching on its luminescence, whilst showing minimal interference from ATP, pyrophosphate and a wide range of biological anions. The probe is equipped with two π-conjugated quinolyl-phenoxyacetate antennae, facilitating excitation at 355 nm in fluorescence microscopy. The ancillary carboxylate groups ensure high water solubility and suppress non-specific binding to albumin protein. Our novel probe demonstrates a level of sensing selectivity for ADP that is unrivaled, producing a linear emission response across the physiologically relevant concentration range (10–400 μM), even in the presence of excess millimolar ATP. We demonstrate that this amphiphilic Eu(III) probe permeates mammalian cells and localises within the mitochondria and lysosomes. The low background emission of the probe combined with its excellent ADP selectivity and long-lived luminescence makes it a promising tool for visualising ADP levels in living cells.