Novel Heterocyclic Analogues of Firefly Luciferin
Novel Heterocyclic Analogues of Firefly Luciferin
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DOI：
10.1021/bi301411d
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发表时间：
2012-12-11
期刊：
影响因子：
2.9
通讯作者：
Branchini, Bruce R.
中科院分区：
文献类型：
--
作者：
Woodroofe, Carolyn C.;Meisenheimer, Poncho L.;Branchini, Bruce R.
合成了以苯并咪唑、苯并呋喃、苯并噻吩、苯并恶唑和吲哚取代天然底物苯并噻唑环体系的五种新型萤火虫荧光素类似物。用重组pyralis Photinus野生型荧光素酶评价了化合物的荧光、生物发光和动力学性质。除了吲哚外，所有含有杂环取代的底物都可以用荧光素酶产生可测量的闪光。与荧光素相比，在不同的最佳pH值和达到最大强度的时间下，反应强度在0.3 ~ 4.4%之间。杂原子的变化对荧光和生物发光发射光谱均有影响，最大发射光谱分别为479 ~ 528 nm和518 ~ 574 nm。虽然在这组结构相似的底物类似物的光谱和生物发光特性方面有一些有趣的趋势，但最重要的发现与含苯并噻吩的化合物有关。这种合成底物产生了缓慢的衰变发光动力学，使荧光素酶的总光基比活性比荧光素值增加了4倍以上。此外，在pH值为6.2-9.4的范围内，发射最大值为523 nm，与天然底物相比，蓝移了37 nm。苯并噻吩荧光素非凡的生物发光特性应该转化为更高的灵敏度，用于各种基于荧光素酶的分析物检测。
Five novel firefly luciferin analogues in which the benzothiazole ring system of the natural substrate was replaced with benzimidazole, benzofuran, benzothiophene, benzoxazole, and indole were synthesized. The fluorescence, bioluminescence, and kinetic properties of the compounds were evaluated with recombinant Photinus pyralis wild type luciferase. With the exception of indole, all of the substrates containing heterocycle substitutions produced readily measurable flashes of light with luciferase. Compared to that of luciferin, the intensities ranged from 0.3 to 4.4% in reactions with varying pH optima and times to reach maximal intensity. The heteroatom changes influenced both the fluorescence and bioluminescence emission spectra, which displayed maxima of 479-528 and 518-574 nm, respectively. While there were some interesting trends in the spectroscopic and bioluminescence properties of this group of structurally similar substrate analogues, the most significant findings were associated with the benzothiophene-containing compound. This synthetic substrate produced slow decay glow kinetics that increased the total light-based specific activity of luciferase more than 4-fold versus the luciferin value. Moreover, over the pH range of 6.2-9.4, the emission maximum is 523 nm, an unusual 37 nm blue shift compared to that of the natural substrate. The extraordinary bioluminescence properties of the benzothiophene luciferin should translate into greater sensitivity for analyte detection in a wide variety of luciferase-based applications.