A biocompatible "split luciferin" reaction and its application for non-invasive bioluminescent imaging of protease activity in living animals.
A biocompatible "split luciferin" reaction and its application for non-invasive bioluminescent imaging of protease activity in living animals.
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DOI：
10.1002/9780470559277.ch140047
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发表时间：
2014-09-09
影响因子：
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通讯作者：
Dubikovskaya, Elena A
中科院分区：
文献类型：
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作者：
Godinat, Aurelien;Budin, Ghyslain;Morales, Alma R;Park, Hyo Min;Sanman, Laura E;Bogyo, Matthew;Yu, Allen;Stahl, Andreas;Dubikovskaya, Elena A
许多人类病理学（例如癌症、糖尿病和神经退行性疾病）非常复杂，需要新的工具来研究整个有机体水平的生物过程。新型生物相容性反应的发现极大地增进了我们对基础生物学的理解，然而，对于在多种人类疾病中发挥非常重要作用的许多人类蛋白酶，目前还没有有效的工具进行实时非侵入性成像。我们最近报道，“分裂荧光素”生物相容性反应是使用生物发光成像（BLI）直接评估活体动物中蛋白酶活性的宝贵工具。由于 BLI 是迄今为止已知的最灵敏的体内成像方式，因此该方法可广泛应用于评估多种蛋白酶活性以及鉴定其新的肽特异性底物。在本协议中，我们描述了这种“分裂荧光素”反应在试管测定和活体动物中定量蛋白酶活性的几种应用。
The great complexity of many human pathologies such as cancer, diabetes, and neurodegenerative diseases requires new tools for studies of biological processes on the whole organism level. The discovery of novel biocompatible reactions has tremendously advanced our understanding of basic biology, however, no efficient tools exist for real-time non-invasive imaging of many human proteases that play very important roles in multiple human disorders. We recently reported that “split luciferin” biocompatible reaction represents a valuable tool for evaluation of protease activity directly in living animals using bioluminescence imaging (BLI). Since BLI is the most sensitive in vivo imaging modality known to date, this method can be widely applied for the evaluation of multiple proteases activity as well as identification of their new peptide-specific substrates. In this protocol we describe several applications of this “split luciferin” reaction for quantification of protease activities in test tube assays and living animals.