Effects of cargo molecules on the cellular uptake of arginine-rich cell-penetrating epeptides
Effects of cargo molecules on the cellular uptake of arginine-rich cell-penetrating epeptides
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DOI：
10.1016/j.bbamem.2005.04.010
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发表时间：
2005-06-30
影响因子：
3.4
通讯作者：
Ottinger, EA
中科院分区：
文献类型：
--
作者：
Maiolo, JR;Ferrer, M;Ottinger, EA
细胞穿透肽（CPP）作为用于细胞内递送缀合分子如肽、蛋白质和寡核苷酸的载体的鉴定已经成为调节细胞内生物活性的重要工具。细胞摄取CPP的机制尚不清楚，根据CPP和细胞类型，似乎既有内吞作用，也有非内吞作用。此外，还不知道货物序列是否对CPP序列的摄取和细胞分布性质具有影响。在这里，我们结合联合收割机的结果，从定量荧光显微镜和结合到脂质膜模型，以确定货物肽分子的细胞摄取和分布的富含精氨酸的CPP，R-7，和R7 W，在活细胞中的效果。使用定量荧光的图像分析算法来测量细胞摄取的肽的相对量，以及摄取本质上是内吞的程度。这里呈现的结果表明，富含精氨酸的CPP与肽序列的融合降低了摄取的效率，并且显著地改变了CPP的细胞分布，从弥散模式变为其中肽大部分保留在内体隔室中的模式。(c)2005 Elsevier B. V.保留所有权利。
The identification of cell-penetrating peptides (CPPs) as vectors for the intracellular delivery of conjugated molecules such as peptides, proteins, and oligonucleotides has emerged as a significant tool to modulate biological activities inside cells. The mechanism of CPP uptake by the cells is still unclear, and appears to be both endocytotic and non-endocytotic, depending on the CPP and cell type. Moreover, it is also unknown whether cargo sequences have an effect on the uptake and Cellular distribution properties of CPP sequences. Here, we combine results from quantitative fluorescence microscopy and binding to lipid membrane models to determine the effect of cargo peptide molecules on the cellular uptake and distribution of the arginine-rich CPPs, R-7, and R7W, in live cells. Image analysis algorithms that quantity fluorescence were used to measure the relative amount of peptide taken up by the cell, as well as the extent to which the uptake was endocytotic in nature. The results presented here indicate that fusion of arginine-rich CPPs to peptide sequences reduces the efficiency of uptake, and dramatically changes the cellular distribution of the CPP from a diffuse pattern to one in which the peptides are mostly retained in endosomal compartments. (c) 2005 Elsevier B.V. All rights reserved.