Covalent Attachment of Cyclic TAT Peptides to GFP Results in Protein Delivery into Live Cells with Immediate Bioavailability
Covalent Attachment of Cyclic TAT Peptides to GFP Results in Protein Delivery into Live Cells with Immediate Bioavailability
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DOI：
10.1002/anie.201410006
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发表时间：
2015-02-02
影响因子：
16.6
通讯作者：
Hackenberger, Christian P. R.
中科院分区：
文献类型：
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作者：
Nischan, Nicole;Herce, Henry D.;Hackenberger, Christian P. R.
通过使用富含精氨酸的细胞穿透肽（CPP）将游离分子递送到细胞质和细胞核中已限于小货物，而大货物如蛋白质被摄取并被捕获在内吞囊泡中。基于最近的工作，其中我们表明，富含精氨酸的CPP的转导效率可以大大提高环化，目的是使用环状CPP运输全长蛋白质，在这项研究中的绿色荧光蛋白（GFP），进入活细胞的胞质溶胶。通过使用叠氮基官能化的CPP和炔官能化的GFP获得环状和线性CPP-GFP缀合物。我们的研究结果表明，环状CPP-GFP缀合物被内化到活细胞中，在胞质溶胶和细胞核中具有立即的生物利用度，而线性CPP类似物不赋予GFP转导。该技术将环状CPP的应用扩展到将功能全长蛋白质有效运输到活细胞中。
The delivery of free molecules into the cytoplasm and nucleus by using arginine-rich cell-penetrating peptides (CPPs) has been limited to small cargoes, while large cargoes such as proteins are taken up and trapped in endocytic vesicles. Based on recent work, in which we showed that the transduction efficiency of arginine-rich CPPs can be greatly enhanced by cyclization, the aim was to use cyclic CPPs to transport full-length proteins, in this study green fluorescent protein (GFP), into the cytosol of living cells. Cyclic and linear CPP-GFP conjugates were obtained by using azido-functionalized CPPs and an alkyne-functionalized GFP. Our findings reveal that the cyclic-CPP-GFP conjugates are internalized into live cells with immediate bioavailability in the cytosol and the nucleus, whereas linear CPP analogues do not confer GFP transduction. This technology expands the application of cyclic CPPs to the efficient transport of functional full-length proteins into live cells.