The Genomics Research Center, Academia Sinica , Taipei 11529, Taiwan.
J Am Chem Soc. 2014 Jul 16;136(28):9953-61. doi: 10.1021/ja5010174. Epub 2014 Jul 3.
We have designed a low fluorescent azido-BODIPY-based probe AzBOCEt (Az10) that undergoes copper(I)-catalyzed 1,3-dipolar cycloadditions with alkynes to yield strongly fluorescent triazole derivatives. The fluorescent quantum yield of a triazole product T10 is enhanced by 52-fold as compared to AzBOCEt upon excitation at a wavelength above 500 nm. Quantum mechanical calculations indicate that the increase in fluorescence upon triazole formation is due to the lowering of the HOMO energy level of the aryl moiety to reduce the process of acceptor photoinduced electron transfer. AzBOCEt is shown to label alkyne-functionalized proteins in vitro and glycoproteins in cells with excellent selectivity, and enables cell imaging and visualization of glycoconjugates in alkynyl-saccharide-treated cells at extremely low concentration (0.1 μM). Furthermore, the alkyne-tagged glycoproteins from cell lysates can be directly detected with AzBOCEt in gel electrophoresis.
我们设计了一种低荧光叠氮基-BODIPY 探针 AzBOCEt（Az10），它可以与炔烃发生铜（I）催化的 1,3-偶极环加成反应，生成强荧光三唑衍生物。与 AzBOCEt 相比，在波长大于 500nm 的激发下，三唑产物 T10 的荧光量子产率增强了 52 倍。量子力学计算表明，三唑形成时荧光的增加是由于芳基部分的 HOMO 能级降低，从而减少了受体光诱导电子转移的过程。AzBOCEt 被证明可以在体外标记炔基化的蛋白质和细胞中的糖蛋白，具有优异的选择性，并能够在极低浓度（0.1μM）下对炔基-糖处理的细胞中的糖缀合物进行细胞成像和可视化。此外，来自细胞裂解物的炔基标记糖蛋白可以在凝胶电泳中直接用 AzBOCEt 检测到。
