Luciferase-Specific Coelenterazine Analogues for Optical Contamination-Free Bioassays.
Luciferase-Specific Coelenterazine Analogues for Optical Contamination-Free Bioassays.
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DOI：
10.1038/s41598-017-00955-6
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发表时间：
2017-04-19
影响因子：
4.6
通讯作者：
Kim SB
中科院分区：
文献类型：
--
作者：
Nishihara R;Abe M;Nishiyama S;Citterio D;Suzuki K;Kim SB
多个光学读数之间的光谱重叠通常会导致荧光和生物发光中的光学污染。为了解决这个问题，我们创建了五种不同的coelenterazine （CTZ）类似物，旨在选择性地照亮具有独特荧光素酶选择性的特定荧光素酶。在实验中，我们发现含有乙基或苯基的CTZ类似物通过修饰CTZ的咪唑吡radinone主链的C-2和C-6位置的官能团，对特定的荧光素酶和ph具有显著的生物发光偏性。荧光素酶特异性CTZ类似物的光学无污染特性得到了例证，该类似物在混合光学探针中阐明了抗雌激素和雷帕霉素的活性。这种独特的生物发光平台具有巨大的潜力，可以在没有光学污染的生物分析中对多个光学读数进行特异性和高通量成像。
Spectral overlaps among the multiple optical readouts commonly cause optical contamination in fluorescence and bioluminescence. To tackle this issue, we created five-different lineages of coelenterazine (CTZ) analogues designed to selectively illuminate a specific luciferase with unique luciferase selectivity. In the attempt, we found that CTZ analogues with ethynyl or styryl groups display dramatically biased bioluminescence to specific luciferases and pHs by modifying the functional groups at the C-2 and C-6 positions of the imidazopyradinone backbone of CTZ. The optical contamination-free feature was exemplified with the luciferase-specific CTZ analogues, which illuminated anti-estrogenic and rapamycin activities in a mixture of optical probes. This unique bioluminescence platform has great potential for specific and high throughput imaging of multiple optical readouts in bioassays without optical contamination.