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首页> 外文期刊>Biochemistry >Targeting of nonkaryophilic cell-permeable peptides into the nuclei of intact cells by covalently attached nuclear localization signals.
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Targeting of nonkaryophilic cell-permeable peptides into the nuclei of intact cells by covalently attached nuclear localization signals.

机译：通过共价附着的核定位信号靶向非核细胞渗透肽进入完整细胞的细胞核。

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摘要

Dermaseptins are a family of antimicrobial peptides that lyse target bacterial cells by destabilization of their membranes. Here we present a novel application of a peptide derived from the dermaseptin S4, S4(13). At nontoxic concentrations, fluorescently labeled S4(13) was able to penetrate intact cultured HeLa cells but essentially failed to enter their nuclei despite its low molecular weight. Covalent attachment of nuclear localization signal (NLS) motifs of the SV40-T-antigen and of the HIV-1 Rev protein (ARM) conferred karyophilic properties upon the S4(13). The resulting peptides, which were designated as PV-S4(13) and RR-S4(13) penetrated into intact HeLa cells and were able to accumulate within the cells' nuclei. In studies with digitonin-permeabilized cells, nuclear uptake of the PV-S4(13) and the RR-S4(13) peptides showed the same features that characterize active nuclear import. Nuclear import was observed at 37 degrees C, was ATP-dependent, and was inhibited by the free peptides bearing the SV40 NLS and the Rev and Tat ARMs. Microinjected S4(13) remained in the cytoplasm while microinjected RR-S4(13) was translocated into the cells' nuclei. The new type of cell-permeable karyophilic lead compound for therapeutic purposes, as a tool to study nucleocytoplasmic shuttling in intact cells, and for the delivery of peptides to the nucleus.
机译：Dermaseptins是一种抗菌肽的家族，其通过搅拌膜来溶解靶细菌细胞。在这里，我们提出了一种衍生自皮塞蛋白S4，S4（13）的肽的新施用。在无毒浓度下，荧光标记的S4（13）能够穿透完整的培养的HeLa细胞，但尽管其分子量低，但基本上未进入它们的核。在S4（13）上赋予S4（13）的核定位信号（NLS）和HIV-1转蛋白（ARM）的核定位信号（NLS）基序的共价附着。将所得肽作为PV-S4（13）和RR-S4（13）指定为完整的HeLa细胞，并能够在细胞的核内积聚。在Digitonin渗透细胞的研究中，PV-S4（13）的核吸收和RR-S4（13）肽显示出具有活跃核导入的相同特征。在37℃下观察到核导入，依赖于ATP依赖性，并被含有SV40 NLS和REV和TAT臂的游离肽抑制。微内注射的S4（13）仍然在细胞质中，而微量注射的RR-S4（13）易于旋转到细胞的细胞核中。用于治疗目的的新型细胞可渗透半核苷酸铅化合物，作为研究完整细胞中的核细胞质穿梭的工具，并用于向细胞核递送肽。

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  • 来源
    《Biochemistry》 |2002年第29期|共7页
  • 作者

    Hariton Gazal E; Feder R; Mor A; Graessmann A; Brack Werner R; Jans D; Gilon C; Loyter A;

  • 作者单位

    Department of Organic Chemistry Institute of Chemistry The Hebrew University of Jerusalem 91904 Jerusalem Israel.;

    Department of Applied Physics and Advanced Technologies (FATA) National Autonomous University of Mexico A.P. 1-1010 Queretaro Qro. 67000 Mexico;

    Department of Applied Physics and Advanced Technologies (FATA) National Autonomous University of Mexico A.P. 1-1010 Queretaro Qro. 67000 Mexico;

    Department of Applied Physics and Advanced Technologies (FATA) National Autonomous University of Mexico A.P. 1-1010 Queretaro Qro. 67000 Mexico;

    Department of Applied Physics and Advanced Technologies (FATA) National Autonomous University of Mexico A.P. 1-1010 Queretaro Qro. 67000 Mexico;

    Department of Applied Physics and Advanced Technologies (FATA) National Autonomous University of Mexico A.P. 1-1010 Queretaro Qro. 67000 Mexico;

    Department of Applied Physics and Advanced Technologies (FATA) National Autonomous University of Mexico A.P. 1-1010 Queretaro Qro. 67000 Mexico;

    Department of Applied Physics and Advanced Technologies (FATA) National Autonomous University of Mexico A.P. 1-1010 Queretaro Qro. 67000 Mexico;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;
  • 关键词

    Peptides; intact cell; Cells; import; 肽类; 细胞;

    机译：Peptides;intact cell;Cells;import;肽类;细胞;

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