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Krysten A Jones¹, William B Porterfield¹, Colin M Rathbun¹

¹Departments of Chemistry, ‡Molecular Biology & Biochemistry, and §Pharmaceutical Sciences, University of California , Irvine, California 92697, United States.

Journal of the American Chemical Society

|January 21, 2017

概括

研究人员通过设计定制的 luciferins 和 luciferases 来开发出新的,可辨别的生物发光探针. 这些设计酶-基质对使复杂细胞网络的多色成像和询问 in vivo 成为可能.

科学领域:

生物化学
分子生物学
生物医学成像

背景情况:

使用 luciferase-luciferin 系统的生物发光成像是生物医学研究中跟踪细胞的关键工具.
目前的系统通常使用相同的基质,限制了在单个受试者中同时区分多种细胞类型的能力.
需要直角探针来实现多色成像和详细的细胞网络分析.

研究的目的:

设计新的可辨别的光酶-光素对,
克服现有探针的局限性, 无法在体内区分多种细胞类型.
开发一个一般的选平台来识别新的酶基质对.

主要方法:

对突变酶库进行反复选,以检测它们是否具有固态修饰的 luciferins.
具有选择性相互作用的正交酶基质对的识别.
在体外和基于细胞的生物发光探测器的验证.

主要成果:

通过工程 luciferases 选择性处理成功制作的定制 luciferins.
在互补作用时产生光的正交酶-基质对.
在体外和培养细胞模型中证明了这些设计对的功能.

结论:

开发的设计型 luciferase-luciferin 双为多组件成像提供可区分的探针.
这些工具可以直接查询细胞网络,扩大当前的体内成像能力.
一般选平台加速了新生物发光工具的发现,增强了这一领域.

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