A Biocompatible in Vivo Ligation Reaction and Its Application for Noninvasive Bioluminescent Imaging of Protease Activity in Living Mice
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- 作者：Aur茅lien Godinat ; Hyo Min Park ; Stephen C. Miller ; Ke Cheng ; Douglas Hanahan ; Laura E. Sanman ; Matthew Bogyo ; Allen Yu ; Gennady F. Nikitin ; Andreas Stahl ; Elena A. Dubikovskaya
- 刊名：ACS Chemical Biology
- 出版年：2013
- 出版时间：May 17, 2013
- 年：2013
- 卷：8
- 期：5
- 页码：987-999
- 全文大小：575K
- 年卷期：v.8,no.5(May 17, 2013)
- ISSN：1554-8937
文摘
The discovery of biocompatible reactions had a tremendous impact on chemical biology, allowing the study of numerous biological processes directly in complex systems. However, despite the fact that multiple biocompatible reactions have been developed in the past decade, very few work well in living mice. Here we report that d-cysteine and 2-cyanobenzothiazoles can selectively react with each other in vivo to generate a luciferin substrate for firefly luciferase. The success of this 鈥硈plit luciferin鈥?ligation reaction has important implications for both in vivo imaging and biocompatible labeling strategies. First, the production of a luciferin substrate can be visualized in a live mouse by bioluminescence imaging (BLI) and furthermore allows interrogation of targeted tissues using a 鈥砪aged鈥?luciferin approach. We therefore applied this reaction to the real-time noninvasive imaging of apoptosis associated with caspase 3/7. Caspase-dependent release of free d-cysteine from the caspase 3/7 peptide substrate Asp-Glu-Val-Asp-d-Cys (DEVD-(d-Cys)) allowed selective reaction with 6-amino-2-cyanobenzothiazole (NH2-CBT) in vivo to form 6-amino-d-luciferin with subsequent light emission from luciferase. Importantly, this strategy was found to be superior to the commercially available DEVD-aminoluciferin substrate for imaging of caspase 3/7 activity. Moreover, the split luciferin approach enables the modular construction of bioluminogenic sensors, where either or both reaction partners could be caged to report on multiple biological events. Lastly, the luciferin ligation reaction is 3 orders of magnitude faster than Staudinger ligation, suggesting further applications for both bioluminescence and specific molecular targeting in vivo.