Cell-penetrating activity of a short-chain ε-poly-l-α-lysine.

Autor: Kaneda K; Graduate School of Bioscience and Biotechnology, Fukui Prefectural University, Eiheiji-cho, Fukui 910-1195, Japan., Takeuchi Y; Graduate School of Bioscience and Biotechnology, Fukui Prefectural University, Eiheiji-cho, Fukui 910-1195, Japan., Yamanaka K; Department of Life Science & Technology, Kansai University, Suita, Osaka 564-8680, Japan., Hasebe F; Graduate School of Bioscience and Biotechnology, Fukui Prefectural University, Eiheiji-cho, Fukui 910-1195, Japan; Fukui Bioincubation Center (FBIC), Fukui Prefectural University, Eiheiji-cho, Fukui 910-1195, Japan., Maruyama C; Graduate School of Bioscience and Biotechnology, Fukui Prefectural University, Eiheiji-cho, Fukui 910-1195, Japan; Fukui Bioincubation Center (FBIC), Fukui Prefectural University, Eiheiji-cho, Fukui 910-1195, Japan. Electronic address: ch-maruyama@g.fpu.ac.jp., Hamano Y; Graduate School of Bioscience and Biotechnology, Fukui Prefectural University, Eiheiji-cho, Fukui 910-1195, Japan; Fukui Bioincubation Center (FBIC), Fukui Prefectural University, Eiheiji-cho, Fukui 910-1195, Japan. Electronic address: y-hamano@g.fpu.ac.jp.
Jazyk: angličtina
Zdroj: Journal of bioscience and bioengineering [J Biosci Bioeng] 2024 Sep; Vol. 138 (3), pp. 249-253. Date of Electronic Publication: 2024 Jul 10.
DOI: 10.1016/j.jbiosc.2024.06.006
Abstrakt: Bacteria produce polycationic homopoly(amino acid)s, which are characterized by isopeptide backbones. We previously demonstrated that two representative bacterial polycationic isopeptides, ε-poly-l-α-lysine consisting of 25-35 l-α-lysine residues (ε-PαL 25-35 ) and ε-poly-l-β-lysine consisting of l-β-lysine residues (ε-PβL 4-13 ), were internalized into mammalian cells by both energy-independent direct penetration and energy-dependent endocytosis/macropinocytosis, and then diffused throughout the cytosol. In this study, we investigated the cell-penetrating activity of an ε-PαL short-chain derivative consisting of 5-14 l-α-lysine residues (ε-PαL 5-14 ) to gain insight into the relationship between the isopeptide-chain length and the manner of cellular internalization. We prepared a conjugate of ε-PαL 5-14 and a fluorescent dye (FAM) by click chemistry, and incubated the resulting polymer, ε-PαL 5-14 -FAM, with HeLa cells. Unlike ε-PαL 25-35 -FAM, ε-PαL 5-14 -FAM was internalized into cells only by energy-dependent endocytosis/macropinocytosis. Furthermore, a high concentration (>50 μM) was required for the internalization events. ε-PαL 5-14 has a chain length almost equal to that of the membrane permeable ε-PβL 4-13 , which can enter cells at low concentrations. Considering that the basicity of the β-amino group is higher than that of α-amino acid at physiological pH, ε-PβL is expected to have a greater cell-penetrating capacity than ε-PαL, provided their isopeptide-chain lengths are similar, suggesting that a more extended chain derivative of ε-PβL would be more advantageous for cellular internalization of cargo proteins than ε-PαL 25-35 .
(Copyright © 2024 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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