Jacek L. Kolanowski

Reversible Fluorescent Probes for Biological Redox States

The redox chemistry of the cell is key to its function and health, and the development of chemical tools to study redox biology is important. While fluxes in oxidative state are essential for healthy cell function, a chronically elevated oxidative capacity is linked to disease. It is therefore essential that probes of biological redox states distinguish between these two conditions by the reversible sensing of changes over time. In this review, we discuss the current progress towards such probes, and identify key directions for future research in this nascent field of vital biological interest.

Mitochondrially targeted fluorescent redox sensors

The balance of oxidants and antioxidants within the cell is crucial for maintaining health, and regulating physiological processes such as signalling. Consequently, imbalances between oxidants and antioxidants are now understood to lead to oxidative stress, a physiological feature that underlies many diseases. These processes have spurred the field of chemical biology to develop a plethora of sensors, both small-molecule and fluorescent protein-based, for the detection of specific oxidizing species and general redox balances within cells. The mitochondrion, in particular, is the site of many vital redox reactions. There is therefore a need to target redox sensors to this particular organelle. It has been well established that targeting mitochondria can be achieved by the use of a lipophilic cation-targeting group, or by utilizing natural peptidic mitochondrial localization sequences. Here, we review how these two approaches have been used by a number of researchers to develop mitochondrially localized fluorescent redox sensors that are already proving useful in providing insights into the roles of reactive oxygen species in the mitochondria.

A Carborane-Containing Fluorophore as a Stain of Cellular Lipid Droplets

The use of fluorescent markers and probes greatly enhances biological investigations but relies on the provision of an array of fluorophores with diverse properties. Herein we report a novel carborane-containing coumarin, 5, which is sufficiently lipophilic to localise in cellular lipid droplets. In non-polar solvents which show comparable polarities to those of a lipid environment, compound 5 exhibits a fluorescence quantum yield two orders of magnitude greater than found in aqueous solvents, adding a further degree of selectivity to lipid droplet imaging. Compound 5 can stain lipid droplets in ex vivo adipocytes as well as in cultured cells, and can be utilised in flow cytometry as well as confocal microscopy.

Reversible magnetogenic cobalt complexes

Cobalt complexes have been extensively used for their catalytic and solid-state magnetic properties, but the solution-state magnetic properties have not yet been widely exploited. Two versatile cobalt ligand scaffolds were investigated for their magnetic properties, which both demonstrated a diamagnetic to paramagnetic transition upon reduction. Notably, one of these complexes, cobalt tris(2-pyridylmethyl)amine (Co-TPA), was capable of cycling between a stable Co(II) species that could induce longitudinal and transverse relaxation of surrounding water protons, and a stable diamagnetic Co(III) species, exhibiting negligible relaxivity effects. Therefore, we propose Co-TPA as a dynamic redox responsive contrast agent for the magnetic resonance imaging (MRI) of hypoxia.

Selective and Reversible Approaches Toward Imaging Redox Signaling Using Small-Molecule Probes

SIGNIFICANCE Recent research has identified key roles for reactive oxygen species (ROS)/reactive nitrogen species (RNS) in redox signaling, but much remains to be uncovered. Molecular imaging tools to study these processes must not only be selective to enable identification of the ROS/RNS involved but also reversible to distinguish signaling processes from oxidative stress. Fluorescent sensors offer the potential to image such processes with high spatial and temporal resolution. RECENT ADVANCES A broad array of strategies has been developed that enable the selective sensing of ROS/RNS. More recently, attention has turned to the design of reversible small-molecule sensors of global redox state, with a further set of probes capable of reversible sensing of individual ROS/RNS. CRITICAL ISSUES In this study, we discuss the key challenges in achieving simultaneous detection of reversible oxidative bursts with unambiguous determination of a particular ROS/RNS. FUTURE DIRECTIONS We have highlighted key design features of small-molecule probes that show promise in enabling the study of redox signaling, identifying essential parameters that must be assessed for any new probe. Antioxid. Redox Signal. 24, 713-730.

A reversible fluorescent probe for monitoring Ag(I) ions

Silver-containing nanomaterials are of interest for their antibiotic properties, for a wide range of applications from medicine to consumer products. However, much remains to be learnt about the degradation of such materials and their effects on human health. While most analyses involve measurement of total silver levels, it is important also to be able to measure concentrations of active free Ag(I) ions. We report here the preparation of a coumarin-based probe, thiocoumarin silver sensor 1 (TcAg1), that responds reversibly to the addition of silver ions through the appearance of a new fluorescence emission peak at 565 nm. Importantly, this peak is not observed in the presence of Hg(II), a common interferent in Ag(I) sensing. To establish the utility of this sensor, we prepared silver-doped phosphate glasses with demonstrated bactericidal properties, and observed the Ag(I) release from these glasses in solutions of different ionic strength. TcAg1 is therefore a useful tool for the study of the environmental and medical effects of silver-containing materials.