Schwartz Edmund C, Saez Lino, Young Michael W, Muir Tom W
Laboratory of Synthetic Protein Chemistry, The Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.
Nat Chem Biol. 2007 Jan;3(1):50-4. doi: 10.1038/nchembio832. Epub 2006 Nov 26.
Control over the timing, location and level of protein activity in vivo is crucial to understanding biological function. Living systems are able to respond to external and internal stimuli rapidly and in a graded fashion by maintaining a pool of proteins whose activities are altered through post-translational modifications. Here we show that the process of protein trans-splicing can be used to modulate enzymatic activity both in cultured cells and in Drosophila melanogaster. We used an optimized conditional protein splicing system to rapidly trigger the in vivo ligation of two inactive fragments of firefly luciferase in a tunable manner. This technique provides a means of controlling enzymatic function with greater speed and precision than with standard genetic techniques and is a useful tool for probing biological processes.
在体内控制蛋白质活性的时间、位置和水平对于理解生物学功能至关重要。生命系统能够通过维持一组蛋白质来快速且以分级方式响应外部和内部刺激，这些蛋白质的活性通过翻译后修饰而改变。在这里，我们表明蛋白质反式剪接过程可用于在培养细胞和黑腹果蝇中调节酶活性。我们使用了优化的条件性蛋白质剪接系统，以可调方式快速触发萤火虫荧光素酶两个无活性片段的体内连接。与标准遗传技术相比，该技术提供了一种以更高速度和精度控制酶功能的方法，并且是探索生物学过程的有用工具。
