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Issue 10, 2014
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Plasma membrane translocation of a protein needle based on a triple-stranded β-helix motif

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Abstract

Plasma membrane translocation is challenging due to the barrier of the cell membrane. Contrary to the synthetic cell-penetrating materials, tailed bacteriophages use cell-puncturing protein needles to puncture the cell membranes as an initial step of the DNA injection process. Cell-puncturing protein needles are thought to remain functional in the native phages. In this paper, we found that a bacteriophage T4 derived protein needle of 16 nm length spontaneously translocates through the living cell membrane. The β-helical protein needle (β-PN) internalizes into human red blood cells that lack endocytic machinery. By comparing the cellular uptake of β-PNs with modified surface charge, it is shown that the uptake efficiency is maximum when it has a negative charge corresponding to a zeta potential value of −16 mV. In HeLa cells, uptake of β-PN incorporates endocytosis independent mechanisms with partial macropinocytosis dependence. The endocytosis dependence of the uptake increases when the surface charges of β-PNs are modified to positive or negative. Thus, these results suggest that natural DNA injecting machinery can serve as an inspiration to design new class of cell-penetrating materials with a tailored mechanism.

Graphical abstract: Plasma membrane translocation of a protein needle based on a triple-stranded β-helix motif

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Supplementary files

Article information


Submitted
14 May 2014
Accepted
25 Jul 2014
First published
01 Aug 2014

Mol. BioSyst., 2014,10, 2677-2683
Article type
Paper

Plasma membrane translocation of a protein needle based on a triple-stranded β-helix motif

N. J. M. Sanghamitra, H. Inaba, F. Arisaka, D. Ohtan Wang, S. Kanamaru, S. Kitagawa and T. Ueno, Mol. BioSyst., 2014, 10, 2677
DOI: 10.1039/C4MB00293H

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