Fluorescence correlation spectroscopy reveals highly efficient cytosolic delivery of certain penta-arg proteins and stapled peptides.
Fluorescence correlation spectroscopy reveals highly efficient cytosolic delivery of certain penta-arg proteins and stapled peptides.
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DOI：
10.1021/ja510391n
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发表时间：
2015-02-25
影响因子：
15
通讯作者：
Schepartz A
中科院分区：
文献类型：
--
作者：
LaRochelle JR;Cobb GB;Steinauer A;Rhoades E;Schepartz A
我们使用荧光相关光谱（FCS），以准确和精确地确定相对效率与三个家庭的“细胞穿透肽”交通到哺乳动物细胞的胞质溶胶。我们发现，某些分子含有“五精氨酸”基序达到胞质溶胶，完整的，效率大于50%。该值比广泛研究的源自HIV达特或聚精氨酸Arg 8的阳离子序列所观察到的值高至少10倍，并且等于在细胞和动物中具有活性的烃钉合肽的值。此外，我们表明，钉合肽到达细胞溶质的效率，如FCS所确定的，与它们在基于细胞的测定中的功效直接相关。我们希望这些发现和相关技术将有助于设计可预测和有效到达哺乳动物细胞内部的肽，蛋白质和肽模拟物。
We used fluorescence correlation spectroscopy (FCS) to accurately and precisely determine the relative efficiencies with which three families of “cell-penetrating peptides” traffic to the cytosol of mammalian cells. We find that certain molecules containing a “penta-arg” motif reach the cytosol, intact, with efficiencies greater than 50%. This value is at least 10-fold higher than that observed for the widely studied cationic sequence derived from HIV Tat or polyarginine Arg8, and equals that of hydrocarbon-stapled peptides that are active in cells and animals. Moreover, we show that the efficiency with which stapled peptides reach the cytosol, as determined by FCS, correlates directly with their efficacy in cell-based assays. We expect that these findings and the associated technology will aid the design of peptides, proteins, and peptide mimetics that predictably and efficiently reach the interior of mammalian cells.