Cargo delivery kinetics of cell-penetrating peptides
Cargo delivery kinetics of cell-penetrating peptides
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DOI：
10.1016/s0005-2736(01)00398-4
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发表时间：
2001-12-01
影响因子：
3.4
通讯作者：
Langel, Ü
中科院分区：
文献类型：
--
作者：
Hällbrink, M;Florén, A;Langel, Ü
细胞穿透肽（CPP）的多样性是已知的，但到目前为止，这些肽的唯一共同点是以能量不依赖的方式获得细胞进入的能力。CPP用于细胞进入的机制在很大程度上是未知的，并且缺乏比较不同肽的数据。为了获得更多关于细胞穿透过程的信息，以及定量比较不同CPP的摄取效率，我们研究了穿透素、转运素、达特（48-60）和MAP（KLAL）的细胞摄取和货物递送动力学。各CPP（用荧光猝灭剂3-硝基酪氨酸标记）通过二硫键与小的五肽货物（用2-氨基苯甲酸荧光团标记）偶联。当CPP-S-S-货物构建体的二硫键在细胞内环境中还原时，货物的细胞摄取记录为荧光强度的增加。我们的数据显示MAP具有最快的摄取，其次是转运蛋白、达特（48-60），最后是穿透蛋白。同样，MAP具有最高的货物递送效率，其次是transportan、达特（48-60），最后是penetratin。由于已经发现一些CPP在高浓度下是有毒的，我们表征了CPP对细胞2-[H-3]脱氧葡萄糖-6-磷酸渗漏的影响。在此系统上的测量表明，膜干扰电位似乎与肽的疏水矩相关。总之，已经定量表征了四种不同CPP的细胞货物递送的产率和动力学。(C)2001 Elsevier Science B. V.保留所有权利。
A diversity of cell-penetrating peptides (CPPs), is known, but so far the only common denominator for these peptides is the ability to gain cell entry in an energy-independent manner. The mechanism used by CPPs for cell entry is largely unknown, and data comparing the different peptides are lacking. In order to gain more information about the cell-penetrating process, as well as to quantitatively compare the uptake efficiency of different CPPs, we have studied the cellular uptake and cargo delivery kinetics of penetratin, transportan, Tat (48-60) and MAP (KLAL). The respective CPPs (labelled with the fluorescence quencher, 3-nitrotyrosine) are coupled to small a pentapeptide cargo (labelled with the 2-amino benzoic acid fluorophore) via a disulfide bond. The cellular uptake of the cargo is registered as an increase in fluorescence intensity when the disulfide bond of the CPP-S-S-cargo construct is reduced in the intracellular milieu. Our data show that MAP has the fastest uptake, followed by transportan, Tat(48-60) and, last, penetratin. Similarly, MAP has the highest cargo delivery efficiency, followed by transportan, Tat (48-60) and, last, penetratin. Since some CPPs have been found to be toxic at high concentration, we characterized the influence of CPPs on cellular 2-[H-3]deoxyglucose-6-phosphate leakage. Measurements on this system show that the membrane-disturbing potential appears to be correlated with the hydrophobic moment of the peptides. In summary, the yield and kinetics of cellular cargo delivery for four different CPPs has been quantitatively characterized. (C) 2001 Elsevier Science B.V. All rights reserved.