Mechanism-Based Design of a Photoactivatable Firefly Luciferase

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• 作者：Jingyi Zhao ; Shixian Lin ; Yong Huang ; Jing Zhao ; Peng R. Chen
• 刊名：The Journal of the American Chemical Society
• 出版年：2013
• 出版时间：May 22, 2013
• 年：2013
• 卷：135
• 期：20
• 页码：7410-7413
• 全文大小：251K
• 年卷期：v.135,no.20(May 22, 2013)
• ISSN：1520-5126

文摘

We developed a photoactivatable firefly luciferase (pfLuc) whose activation can be controlled by light. A photocaged Lys analogue was site-specifically incorporated into fLuc to replace its key catalytic Lys residue, Lys529, rendering fLuc inactive until light-triggered removal of the caging group. This photoinduced gain of luminescence provides a facile approach for assessing the photolysis efficiency of this valuable photosensitive Lys analogue within the context of its carrier protein in vitro and in living cells. We further took advantage of the spatial and temporal activation feature of pfLuc for intracellular measurement of labile ATP levels without impairment of cellular physiology.