Yuming Yang,†,§ Qiang Zhao,‡,§ Wei Feng,† and Fuyou Li*,† †Department of Chemistry and State Key Laboratory of Molecular Engineering of Polymers and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, P. R. China ‡Key Laboratory for Organic Electronics and Information Displays (KLOEID) and Institute of Advanced Materials (IAM), Nanjing University of Posts and Telecommunications, Nanjing 210046, P. R. China.

Luminescent chemodosimeters for bioimaging.

Design strategies for water-soluble small molecular chromogenic and fluorogenic probes.

Click Chemistry for Drug Development and Diverse Chemical–Biology Applications

Due to the biological importances of thiols, such as cysteine, homocysteine and glutathione, the development of optical probes for thiols has been an active research area in recent few years. This critical review focuses on the fluorescent or colorimetric sensors for thiols according to their unique mechanisms between sensors and thiols, including Michael addition, cyclization with aldehyde, cleavage of sulfonamide and sulfonate ester by thiols, cleavage of selenium–nitrogen bond by thiols, cleavage of disulfide by thiols, metal complexes-oxidation–reduction, metal complexes-displace coordination, nano-particles and others (110 references).

Fluorescent and colorimetric probes for detection of thiols

Microscopy and mass spectrometry (MS) are complementary techniques: The former provides spatiotemporal information in living cells, but only for a handful of recombinant proteins at a time, whereas the latter can detect thousands of endogenous proteins simultaneously, but only in lysed samples. Here, we introduce technology that combines these strengths by offering spatially and temporally resolved proteomic maps of endogenous proteins within living cells. Our method relies on a genetically targetable peroxidase enzyme that biotinylates nearby proteins, which are subsequently purified and identified by MS. We used this approach to identify 495 proteins within the human mitochondrial matrix, including 31 not previously linked to mitochondria. The labeling was exceptionally specific and distinguished between inner membrane proteins facing the matrix versus the intermembrane space (IMS). Several proteins previously thought to reside in the IMS or outer membrane, including protoporphyrinogen oxidase, were reassigned to the matrix by our proteomic data and confirmed by electron microscopy. The specificity of peroxidase-mediated proteomic mapping in live cells, combined with its ease of use, offers biologists a powerful tool for understanding the molecular composition of living cells.

/pdf/proteomic-mapping-of-mitochondria-in-living-cells-via-2e93umv0r1.pdf

Proteomic Mapping of Mitochondria in Living Cells via Spatially Restricted Enzymatic Tagging

Fast detection of cellular thiols in aqueous medium was achieved using a newly developed fluorescence probe (see picture). Based on this probe, a high-throughput fluorescence assay for glutathione reductase was developed.

A Highly Sensitive Fluorescence Probe for Fast Thiol-Quantification Assay of Glutathione Reductase†

Near-infrared (NIR) fluorescence-based sensors capable of selective detection of H2S in vivo would be useful tools to understand the mechanisms of diseases. A new NIR fluorescence probe 1 was developed for the detection of endogenous H2S in colorectal cancer cells in mice. 1 displayed an 87-fold fluorescence enhancement at 796 nm (with excitation at 730 nm) when reacted with H2S in a buffer (pH 7.4). 1 was water-soluble, cell-membrane-permeable, had low cytotoxicity and high selectivity and sensitivity for H2S. The properties of 1 enable its use in monitoring endogenous H2S in living cells, tissues, and mice. The bioimaging results indicated that (1) D-Cys could induce endogenous H2S production in living cells and stimulate angiogenesis; (2) tail intravenous injection of 1 into mice generated strong fluorescence in the liver while intraperitoneal injection of D-Cys could further enhance fluorescence in the liver in vivo; (3) importantly, endogenous H2S in colorectal cancer cells (HCT116, HT29) in vitro and in murine tumor models could be quickly and selectively detected by intratumoral injection of 1. These results indicated that our new probe could serve as an efficient tool for the detection of cellular H2S in living animals and even for cancer diagnosis.

/pdf/a-new-h2s-specific-near-infrared-fluorescence-enhanced-probe-4j97sid12x.pdf

A new H2S-specific near-infrared fluorescence-enhanced probe that can visualize the H2S level in colorectal cancer cells in mice.

Hydrogen sulfide (H2S) is an important endogenous signalling molecule and also an important environmental target for detection. New reaction-based colorimetric and fluorescent turn-on probes based on selective thiolyling of NBD (7-nitro-1,2,3-benzoxadiazole) ether were explored for sensing of H2S in aqueous buffer. The syntheses of both probes are simple and quite straightforward. The probes are highly sensitive and selective toward H2S over other biologically relevant species. Probe 1 can be used to directly visualize H2S by the naked eye and shows more than 1000-fold fluorescence increase upon reaction with H2S. Probe 2 is a near-infrared fluorescent sensor for H2S at physiological pH.

Zhen Xi

Papers

A Highly Sensitive Fluorescence Probe for Fast Thiol-Quantification Assay of Glutathione Reductase†

A new H2S-specific near-infrared fluorescence-enhanced probe that can visualize the H2S level in colorectal cancer cells in mice.

NBD-based colorimetric and fluorescent turn-on probes for hydrogen sulfide

A FRET–ICT dual-quenching fluorescent probe with large off–on response for H2S: synthesis, spectra and bioimaging

A colorimetric and “turn-on” fluorescent chemosensor for Zn(II) based on coumarin Shiff-base derivative