Development of Luciferin Analogues Bearing an Amino Group and Their Application as BRET Donors
Development of Luciferin Analogues Bearing an Amino Group and Their Application as BRET Donors
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DOI：
10.1002/asia.201000219
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发表时间：
2010-09-03
影响因子：
4.1
通讯作者：
Nagano, Tetsuo
中科院分区：
文献类型：
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作者：
Takakura, Hideo;Sasakura, Kiyoshi;Nagano, Tetsuo
我们系统地合成了在苯并噻唑、喹啉、萘和香豆素支架上带有氨基的生物发光底物。它们发出各种颜色的生物发光：红色、橙色、黄色和绿色。氨基取代的香豆基荧光素衍生物香豆基氨基荧光素（CAL）在迄今为止报道的底物中显示出最短的生物发光波长。此外，CAL 的荧光不表现出溶剂化显色现象，这表明其生物发光不易受到环境因素的影响。我们应用CAL作为生物发光共振能量转移（BRET）系统的能量供体底物，以萤火虫荧光素酶突变体点击甲虫红色荧光素酶（CBRluc）作为能量供体酶，黄色荧光蛋白（YFP）作为能量受体荧光团，并获得了清晰的双峰生物发光光谱。对于生物测定中的可靠测量来说，非常需要不受环境因素影响的稳定生物发光。
We systematically synthesized bioluminogenic substrates bearing an amino group on benzothiazole, quinoline, naphthalene, and coumarin scaffolds. They emit bioluminescence in various colors: red, orange, yellow, and green. An amino-substituted coumarylluciferin derivative, coumarylaminoluciferin (CAL), showed the shortest bioluminescence wavelength among substrates reported so far. Further, the fluorescence of CAL did not exhibit solvatochromism, which suggests that its bioluminescence is not susceptible to environmental factors. We applied CAL as an energy-donor substrate for a bioluminescence resonance energy transfer (BRET) system with click beetle red luciferase (CBRluc), a mutant of firefly luciferase, as the energy-donor enzyme and yellow fluorescent protein (YFP) as the energy-acceptor fluorophore, and obtained a clearly bimodal bioluminescence spectrum. Stable bioluminescence that is not influenced by environmental factors is highly desirable for reliable measurements in biological assays.