[7‐(Dialkylamino)coumarin‐4‐yl]methyl‐Caged Compounds as Ultrafast and Effective Long‐Wavelength Phototriggers of 8Bromo‐Substituted Cyclic Nucleotides

Abstract

[7-(Dimethylamino)coumarin-4-yl]methyl (DMACM) and [7-(diethylamino)coumarin-4-yl]methyl (DEACM) esters of 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP) and 8-bromoguanosine 3',5'-cyclic monophosphate (8-Br-cGMP) are described as novel caged compounds for 8-bromo-substituted cyclic nucleotides. Synthesis is accomplished by treatment of the free acids of the cyclic nucleotides with the corresponding 7(dialkylamino)-substituted 4(diazomethyl)coumarins. Irradiation of the DMACM- and DEACM-caged cyclic nucleotides with UV light stimulates the release of the cyclic nucleotides within roughly a nanosecond. The new caged compounds are resistant to hydrolysis in aqueous buffers and exhibit long-wavelength absorption properties with maxima at 400 nm, high extinction coefficients, and high quantum yields (0.15-0.31). Their favorable properties render these compounds the most efficient and rapid phototriggers of 8-bromo-substituted cyclic nucleotides known. The usefulness of the compounds for physiological studies under nondamaging light conditions was examined in HEK293 cells expressing the alpha subunit of the cyclic-nucleotide-gated (CNG) channel of cone photoreceptors (CNGA3) and of olfactory neurons (CNGA2) by using confocal laser scanning microscopy and the patch clamp technique.

(Coumarin‐4‐yl)methyl Esters as Highly Efficient, Ultrafast Phototriggers for Protons and Their Application to Acidifying Membrane Surfaces

“…Both the DMCM and the DMACM moieties have been introduced earlier for the photoreversible inactivation of cyclic nucleotide monophosphates. [13,14] The analogues (7-methoxycoumarin-4-yl)methyl (MCM) diethyl phosphate and methanesulfonate have been described previously, [15,16] but they were not classified or used as phototriggers for protons.…”

(Cumarin‐4‐yl)methylester als hocheffiziente und ultraschnelle Phototrigger für Protonen und ihre Anwendung zur Oberflächenansäuerung von Membranen

“…Die DMCM-und die DMACM-Chromophore wurden für die photoreversible Inaktivierung cyclischer Nucleosidmonophosphate eingeführt. [13,14] Die Analoga (7-Methoxycumarin-4-yl)methyl(MCM)-Diethylphosphat und MCM-Methansulfonat wurden schon früher beschrieben, [15,16] aber nicht als Phototrigger für Protonen klassifiziert oder eingesetzt.…”

Intracellular photoactivation of caged cGMP induces myosin II and actin responses in motile cells

“…This can be seen in Fig. 3C, where the corresponding time traces for the ratio of cortical and cytosolic 30 Configurations for (B) placing the uncaging laser outside and (C) placing the uncaging laser inside the cell. Note that although the laser is focused to an uncaging region of a circle with 1.071 mm (21 pixels) diameter, stray light can still cause uncaging inside neighboring cells.…”