An improved synthesis of dansylated α-galactosylceramide and its use as a fluorescent probe for the monitoring of glycolipid uptake by cells

Cheng, Janice M. H.; Chee, Stephanie H.; Knight, Deborah; Acha‐Orbea, Hans; Hermans, Ian F.; Timmer, Mattie S. M.; Stocker, Bridget L.

doi:10.1016/j.carres.2011.02.014

Cited by 32 publications

(25 citation statements)

References 69 publications

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“…45 Several fluorescent compounds have been designed as tools for monitoring the cellu-lar entry of e.g. 56 7-Nitro-2,1,3benzoxadiazole (NBD) and dansyl served as particularly used fluorophores, 46,[49][50][51]54,57 but also fluorescein, 48,52 indocarbocyanines, 53 rhenium(I) complexes, 56 or BODIPYs have been applied. 56 7-Nitro-2,1,3benzoxadiazole (NBD) and dansyl served as particularly used fluorophores, 46,[49][50][51]54,57 but also fluorescein, 48,52 indocarbocyanines, 53 rhenium(I) complexes, 56 or BODIPYs have been applied.…”

Section: Resultsmentioning

confidence: 99%

Design, characterization and cellular uptake studies of fluorescence-labeled prototypic cathepsin inhibitors

et al. 2015

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Besides their extracellular activity crucial for several pathophysiological conditions, human cysteine cathepsins, in particular cathepsins K and S, represent important intracellular targets for drug development. In the present study, a prototypic dipeptide nitrile inhibitor structure was equipped with a coumarin moiety to function as a fluorescent reporter group. In a second inhibitor, a PEG linker was introduced between the dipeptide nitrile and the fluorophore. These tool compounds 6 and 7 were characterized by kinetic investigations as covalent reversible inhibitors of human cathepsins L, S, K and B. Probe 6 showed a pronounced inhibitory activity against cathepsins K and S, which was corroborated by modeling of inhibition modes. Probe 7 was highly potent (Ki = 93 nM) and selective for cathepsin S. To examine the ability of both probes to enter living cells, human embryonic kidney 293 cells were targeted. At a concentration of 10 μM, cellular uptake of probe 6 was demonstrated by fluorescence measurement after an incubation time of 30 min and 3 h, respectively. The probe's concentration in cell lysates was ascertained on the basis of the emission at 492 nm upon excitation at 450 nm, and the results were expressed as concentrations of probe 6 relative to the protein concentration originating from the lysate. After incubation of 10 μM of probe 6 for 3 h, the cellular uptake was confirmed by fluorescence microscopy. HPLC was used to assess the probes’ lipophilicity, and the obtained

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Section: Resultsmentioning

confidence: 99%

Design, characterization and cellular uptake studies of fluorescence-labeled prototypic cathepsin inhibitors

et al. 2015

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“…These analogs further provided evidence that the 6″-hydroxyl position was not necessary for iNKT cell stimulation, since they stimulated iNKT cells comparably to α-GalCer. Xia et al [114] and Cheng et al [115] reported flow cytometry data for the biotin and dansyl labeled analogs, respectively. These reports demonstrated that these 6″-α-GalCer analogs can be used as probes for the analysis of glycolipid uptake.…”

Section: α-Galcer Analogsmentioning

confidence: 99%

Stimulation of Natural Killer T Cells by Glycolipids

et al. 2013

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Natural killer T (NKT) cells are a subset of T cells that recognize glycolipid antigens presented by the CD1d protein. The initial discovery of immunostimulatory glycolipids from a marine sponge and the T cells that respond to the compounds has led to extensive research by chemists and immunologists to understand how glycolipids are recognized, possible responses by NKT cells, and the structural features of glycolipids necessary for stimulatory activity. The presence of this cell type in humans and most mammals suggests that it plays critical roles in antigen recognition and the interface between innate and adaptive immunity. Both endogenous and exogenous natural antigens for NKT cells have been identified, and it is likely that glycolipid antigens remain to be discovered. Multiple series of structurally varied glycolipids have been synthesized and tested for stimulatory activity. The structural features of glycolipids necessary for NKT cell stimulation are moderately well understood, and designed compounds have proven to be much more potent antigens than their natural counterparts. Nevertheless, control over NKT cell responses by designed glycolipids has not been optimized, and further research will be required to fully reveal the therapeutic potential of this cell type.

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“…For details of the synthesis, see Cheng et al (2011). For related structures, see Sikorski et al (2009), Robertson & Sheldrick (1965), Zhou et al (2002), Kurhade et al(2011).…”

Section: Related Literaturementioning

confidence: 99%

Methyl 6-azido-6-deoxy-α-D-galactoside

Cheng

Gulab²,

Timmer

et al. 2011

Acta Cryst E

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The structure of the title compound, C7H13N3O5, was solved using data from a multiple fragment crystal. The galactoside ring adopts a 4 C 1 chair conformation. In the crystal, the molecules are linked by strong O—H⋯O hydrogen bonds, which build linkages around the screw axis of the cell in a similar way to the iodo analogue. These C-5 and C-6 packing motifs expand to R 2 2(10), C 2 2(7) and C 2 2 2(8) motifs, as found in closely related compounds.

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An improved synthesis of dansylated α-galactosylceramide and its use as a fluorescent probe for the monitoring of glycolipid uptake by cells

Cited by 32 publications

References 69 publications

Design, characterization and cellular uptake studies of fluorescence-labeled prototypic cathepsin inhibitors

Design, characterization and cellular uptake studies of fluorescence-labeled prototypic cathepsin inhibitors

Stimulation of Natural Killer T Cells by Glycolipids

Methyl 6-azido-6-deoxy-α-D-galactoside

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