Design and synthesis of an enzyme activity-based labeling molecule with fluorescence spectral change
Design and synthesis of an enzyme activity-based labeling molecule with fluorescence spectral change
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DOI：
10.1021/ja0657307
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发表时间：
2006-12-20
影响因子：
15
通讯作者：
Nagano, Tetsuo
中科院分区：
文献类型：
--
作者：
Komatsu, Toru;Kikuchi, Kazuya;Nagano, Tetsuo
用小分子染料共价标记特定的标签蛋白质的方法在研究活细胞中蛋白质的动力学行为中起着重要的作用。在醌甲基化化学的基础上，我们设计并合成了一种β-半乳糖苷酶标记探针CMFβ-gal，该探针在标记过程中，由于荧光共振能量转移（FRET）的作用，其荧光波长会发生变化。由于FRET效率随着标记反应而变化，因此标记蛋白质的荧光可以与未反应的探针的荧光分开观察，因此可以立即检测目标蛋白质。这是第一个报告的蛋白质标记探针，其特征是在反应时荧光波长的变化，允许标记的蛋白质被检测，即使在未反应的探针的存在下。
Methods of covalent labeling of a specific tag protein with small-molecular dyes play an important role in studying dynamic behaviors of proteins in living cells. On the basis of quinone methide chemistry, we designed and synthesized a β-galactosidase labeling probe, CMFβ-gal, which shows a fluorescence wavelength change accompanying the labeling reaction, owing to fluorescence resonance energy transfer (FRET). Since the FRET efficiency changes accompanying the labeling reaction, fluorescence of labeled protein can be observed separately from that of the unreacted probe, so immediate detection of the target protein is possible. This is the first report of a protein labeling probe which features a change of fluorescence wavelength upon reaction, allowing the labeled protein to be detected even in the presence of unreacted probe.