Bhc-cNMPs as either water-soluble or membrane-permeant photoreleasable cyclic nucleotides for both one- and two-photon excitation
Bhc-cNMPs as either water-soluble or membrane-permeant photoreleasable cyclic nucleotides for both one- and two-photon excitation
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DOI：
10.1002/cbic.200300814
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发表时间：
2004-08-06
期刊：
影响因子：
3.2
通讯作者：
Tsien, RY
中科院分区：
文献类型：
--
作者：
Furuta, T;Takeuchi, H;Tsien, RY
环状单磷酸核苷 (cNMP) 在许多细胞调节过程中发挥着关键作用，例如生长、分化、运动和基因表达。可以通过照射激活的笼状衍生物可能是研究细胞内第二信使的多种功能的有力工具，因为这些分子的时空动力学可以通过适当聚焦的光照射来控制。在这里，我们报告了作为新的笼状第二信使的 6-溴-7-羟基香豆素-4-基甲酯 (Bhc-cNMP) 及其乙酰基衍生物 (Bhc-cNMP/Ac) 的合成、光化学和生物测试。 Bhc-cNMP 的辐照定量产生了母体 cNMP，其单光子解笼效率 (Phiepsilon) 比 2-硝基苯乙基 (NPE) cNMP 的效率高出一个数量级。此外，可以观察到 Bhc-cNMP（7 和 8）中双光子诱导的 cNMP 光化学释放，双光子解笼作用截面（delta(u)）高达 2.28 GM（1 GM = 10(-50) cm(4) s photon(-1)），这是已报道的 Bhc 笼化合物中的最大值。 δ(u) 值 7 的波长依赖性表明峰值波长是单光子吸收最大值的两倍。 Bhc-cNMPs 显示出实用的水溶性（接近 500 μm），而 7-乙酰化衍生物 (Bhc-cNMPs/Ac) 预计具有一定的膜渗透性。它们的 1 优势在两种类型的生物系统中得到了证明：在蝾螈嗅觉受体细胞中打开 cAMP 介导的转导通道，以及在青鳉鱼鳞片的表皮黑素细胞中 cAMP 介导的运动反应。两个例子都表明 Bhc 和 Bhc/Ac 笼状化合物在许多细胞生物学应用中具有巨大的潜力。
Cyclic nucleoside monophosphates (cNMPs) play key roles in many cellular regulatory processes, such as growth, differentiation, motility, and gene expression. Caged derivatives that can be activated by irradiation could be powerful tools for studying such diverse functions of intracellular second messengers, since the spatiotemporal dynamics of these molecules con be controlled by irradiation with appropriately focused light. Here we report the synthesis, photochemistry, and biological testing of 6-bromo-7-hydroxycoumarin-4-ylmethyl esters of cNMP (Bhc-cNMP) and their acetyl derivatives (Bhc-cNMP/Ac) as new caged second messengers. Irradiation of Bhc-cNMPs quantitatively produced the parent cNMPs with one-photon uncaging efficiencies (Phiepsilon) of up to one order of magnitude better than those of 2-nitrophenethyl (NPE) cNMPs. In addition, two-photon induced photochemical release of cNMP from Bhc-cNMPs (7 and 8) con be observed with the two-photon uncaging action cross-sections (delta(u)) of up to 2.28 GM (1 GM = 10(-50) cm(4) s photon(-1)), which is the largest value among those of the reported Bhc-caged compounds. The wavelength dependence of the delta(u) values of 7 revealed that the peak wavelength was twice that of the one-photon absorption maximum. Bhc-cNMPs showed practically useful water solubility (nearly 500 mum), whereas 7-acetylated derivatives (Bhc-cNMPs/Ac) were expected to have a certain membrane permeability. Their 1 advantages were demonstrated in two types of biological Systems: the opening of cAMP-mediated transduction channels in newt olfactory receptor cells and cAMP-mediated motility responses in epidermal melanophores in scales from medaka fish. I Both examples showed that Bhc and Bhc/Ac caged compounds have great potential for use in many cell biological applications.