...rhodamines as scaffolds for high-contrast fluorogenic probes

来自 : 发布时间：2024-12-22

ActionsCite Favorites Display options Display options Format Carbofluoresceins and carborhodamines as scaffolds for high-contrast fluorogenic probes 1 Janelia Farm Research Campus, Howard Hughes Medical Institute , Ashburn, Virginia, 20147, United States. Carbofluoresceins and carborhodamines as scaffolds for high-contrast fluorogenic probes 1 Janelia Farm Research Campus, Howard Hughes Medical Institute , Ashburn, Virginia, 20147, United States. Fluorogenic molecules are important tools for advanced biochemical and biological experiments. The extant collection of fluorogenic probes is incomplete, however, leaving regions of the electromagnetic spectrum unutilized. Here, we synthesize green-excited fluorescent and fluorogenic analogues of the classic fluorescein and rhodamine 110 fluorophores by replacement of the xanthene oxygen with a quaternary carbon. These anthracenyl \"carbofluorescein\" and \"carborhodamine 110\" fluorophores exhibit excellent fluorescent properties and can be masked with enzyme- and photolabile groups to prepare high-contrast fluorogenic molecules useful for live cell imaging experiments and super-resolution microscopy. Our divergent approach to these red-shifted dye scaffolds will enable the preparation of numerous novel fluorogenic probes with high biological utility. (a) Chemical structures of xanthene dyes and carbon-containing isologues. (b) Existing synthetic strategy… (a) Chemical structures of xanthene dyes and carbon-containingisologues. (b) Existing synthetic strategy to carborhodamine dyes.(c) Divergent synthesis of carborhodamines through carbofluoresceinintermediates.Properties of fluorescent dyes. (a) Spectral properties of dyes. (b) Normalized absorbance at… dyes. (a) Spectral properties of dyes.(b) Normalized absorbance at λmax versus pH for fluorescein(1) and carbofluorescein (4). Error barsshow standard error (SE; n = 2). Determined values(±SE): compound 1, pKa = 6.35 ± 0.02, Hill coefficient = 0.93 ± 0.02; compound 4, pKa = 7.44 ± 0.01, Hillcoefficient = 1.33 ± 0.03. (c) Absorption at λmax versus dielectric constant for rhodamine 110 (2) andcarborhodamine 110 (5). Error bars show SE (n = 2).Confocal microscopy of live, unwashed HeLa cells incubated with esterase substrates 20 or… live, unwashed HeLa cells incubated withesterase substrates 20 or 26 and counterstainedwith Hoechst 33342; scale bars = 10 μm. (a) Compound 20, 1 h incubation. (b) Compound 26, 24 h incubation.PALM imaging of F-actin in fixed mouse embryonic fibroblasts. (a) Synthesis of caged… of F-actin in fixed mouse embryonic fibroblasts. (a)Synthesis of caged carborhodamine 110–phalloidin conjugate 32. Reagents and conditions: (a) TFA/CH2Cl2, rt, 98%. (b) TSTU, DIEA, DMF, 74%. (c) i. H2N-PEG8-CO2H, DIEA, DMF; ii. TSTU, DIEA, DMF, then phalloidin-NH2, 69%, two steps. (b, c) PALM images for labeling with CRh110–PEG8–phallodin conjugate 32. (d, e) Labeling with mEos2–actin. (f, g) Labelingwith AF647–phalloidin. Images are rendered as a heat map oflocalization probability/nm2 with scale in lower right.For clarity, PALM images were plotted at a minimum localization precisionof 22 nm (b, d, and f) and 11 nm (c, e, and g). Scale bars (lowerleft): 5 μm (b, d, and f) or 1 μm (c, e, and g). Zheng H, et al. Chem Commun (Camb). 2013 Jan 18;49(5):429-47. doi: 10.1039/c2cc35997a. Chem Commun (Camb). 2013. PMID: 23164947 Zhang X, et al. Chemistry. 2021 Feb 19;27(11):3688-3693. doi: 10.1002/chem.202005296. Epub 2021 Jan 26. Chemistry. 2021. PMID: 33330995 Zhou X, et al. Angew Chem Int Ed Engl. 2017 Apr 3;56(15):4197-4200. doi: 10.1002/anie.201612628. Epub 2017 Mar 20. Angew Chem Int Ed Engl. 2017. PMID: 28319304 Free PMC article. Yakugaku Zasshi. 2019;139(2):277-283. doi: 10.1248/yakushi.18-00174-3. Yakugaku Zasshi. 2019. PMID: 30713240 Lavis LD. 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