Measuring In Vivo Protein Half-Life
Measuring In Vivo Protein Half-Life
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DOI：
10.1016/j.chembiol.2011.03.014
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发表时间：
2011-06-24
影响因子：
--
通讯作者：
Trono, Didier
中科院分区：
文献类型：
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作者：
Bojkowska, Karolina;de Sio, Francesca Santoni;Trono, Didier
蛋白质周转严重影响许多生物学功能，但一直缺乏方法来评估这个参数在体内。在这里，我们演示了如何SNAP标签融合蛋白的化学标记可以利用来测量在活小鼠中的居民细胞内和细胞外蛋白的半衰期。首先，我们证明了SNAP标签底物在小鼠中具有广泛的生物利用度，可用于特异性的SNAP标签融合蛋白的体内标记。然后，我们应用近红外探针对体内标记的肿瘤进行非侵入性成像。最后，我们使用SNAP介导的化学脉冲追踪标记来测量不同细胞外和细胞内蛋白质的体内半衰期。这些结果为研究活体动物的蛋白质功能开辟了广阔的前景。
Protein turnover critically influences many biological functions, yet methods have been lacking to assess this parameter in vivo. Here, we demonstrate how chemical labeling of SNAP-tag fusion proteins can be exploited to measure the half-life of resident intracellular and extracellular proteins in living mice. First, we demonstrate that SNAP-tag substrates have wide bioavailability in mice and can be used for the specific in vivo labeling of SNAP-tag fusion proteins. We then apply near-infrared probes to perform noninvasive imaging of in vivo-labeled tumors. Finally, we use SNAP-mediated chemical pulse-chase labeling to perform measurement of the in vivo half-life of different extra- and intracellular proteins. These results open broad perspectives for studying protein function in living animals.