Suzuki Tomoki, Futaki Shiroh, Niwa Miki, Tanaka Seigo, Ueda Kunihiro, Sugiura Yukio
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
J Biol Chem. 2002 Jan 25;277(4):2437-43. doi: 10.1074/jbc.M110017200. Epub 2001 Nov 15.
Basic peptides such as human immunodeficiency virus type 1 (HIV-1) Tat-(48-60) and Drosophila Antennapedia-(43-58) have been reported to have a membrane permeability and a carrier function for intracellular protein delivery. We have shown that not only Tat-(48-60) but many arginine-rich peptides, including HIV-1 Rev-(34-50) and octaarginine (Arg(8)), efficiently translocated through the cell membranes and worked as protein carriers (Futaki, S., Suzuki, T., Ohashi, W., Yagami, T., Tanaka, S., Ueda, K., and Sugiura, Y. (2001) J. Biol. Chem. 276, 5836-5840). Quantification and time course analyses of the cellular uptake of the above peptides by mouse macrophage RAW264.7, human cervical carcinoma HeLa, and simian kidney COS-7 cells revealed that Rev-(34-50) and Arg(8) had a comparable translocation efficiency to Tat-(48-60). Internalization of Tat-(48-60) and Rev-(34-50) was saturable and inhibited by the excess addition of the other peptide. Typical endocytosis and metabolic inhibitors had little effect on the internalization. The uptake of these peptides was significantly inhibited in the presence of heparan sulfate or chondroitin sulfates A, B, and C. Treatment of the cells with the anti-heparan sulfate antibody or heparinase III also lowered the translocation of these peptides. These results strongly suggest that the arginine-rich basic peptides share a certain part of the internalization pathway.
据报道，诸如人类免疫缺陷病毒1型（HIV-1）Tat-（48-60）和果蝇触角足蛋白-（43-58）等碱性肽具有膜通透性以及用于细胞内蛋白质递送的载体功能。我们已经表明，不仅Tat-（48-60），而且许多富含精氨酸的肽，包括HIV-1 Rev-（34-50）和八聚精氨酸（Arg(8)），都能有效地穿过细胞膜并作为蛋白质载体发挥作用（Futaki, S., Suzuki, T., Ohashi, W., Yagami, T., Tanaka, S., Ueda, K., and Sugiura, Y. (2001) J. Biol. Chem. 276, 5836-5840）。对小鼠巨噬细胞RAW264.7、人宫颈癌HeLa细胞和猴肾COS-7细胞摄取上述肽的定量和时间进程分析表明，Rev-（34-50）和Arg(8)具有与Tat-（48-60）相当的转运效率。Tat-（48-60）和Rev-（34-50）的内化是可饱和的，并且过量添加另一种肽会抑制其内化。典型的内吞作用和代谢抑制剂对其内化几乎没有影响。在硫酸乙酰肝素或硫酸软骨素A、B和C存在的情况下，这些肽的摄取受到显著抑制。用抗硫酸乙酰肝素抗体或肝素酶III处理细胞也会降低这些肽的转运。这些结果强烈表明，富含精氨酸的碱性肽共享内化途径的某一部分。
