Superluminescent Variants of Marine Luciferases for Bioassays
Superluminescent Variants of Marine Luciferases for Bioassays
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DOI：
10.1021/ac2021882
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发表时间：
2011-11-15
影响因子：
7.4
通讯作者：
Tao, Hiroaki
中科院分区：
文献类型：
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作者：
Kim, Sung Bae;Suzuki, Hideyuki;Tao, Hiroaki
在这项研究中，一个合理的合成超发光的变种，从海洋生物发光酶延长已被证明。一个假定的活性位点的模式海洋荧光素酶，高斯princeps荧光素酶（GLuc），分配，并通过定点诱变修饰。发现有效的变体产生高达10倍强的生物发光，与天然GLuc相比，天然腔肠素发射高达33 nm的红移，在生物测定中呈现有效的光学特征。用哺乳动物双杂交试验、单链探针和BALB/c裸鼠中小鼠B16黑色素瘤转移证明了有利的性质。工程GLuc的独特想法被证明是有效的，即使是其他海洋生物。
In this study, a rational synthesis of superluminescent variants from marine luciferases with prolonged bioluminescence has been demonstrated. A putative active site of a model marine luciferase, Gaussia princeps Luciferase (GLuc), was assigned, and modified by a site-directed mutagenesis. The potent variants were found to generate up to 10 times stronger bioluminescence, emitting red shifts of up to 33 nm with natural coelenterazine than native GLuc, rendering an efficient optical signature in bioassays. The advantageous properties were demonstrated with mammalian two-hybrid assays, single-chain probes,, and metastases of murine B16 melanoma in BALB/c nude mice. The unique ideas for engineering GLuc are proved to be valid even for other marine luciferases.