Comparative Mechanisms of Protein Transduction Mediated by Cell-Penetrating Peptides in Prokaryotes

Abstract

Bacterial and archaeal cell envelopes are complex multilayered barriers that serve to protect these microorganisms from their extremely harsh and often hostile environments. Import of exogenous proteins and nanoparticles into cells is important for biotechnological applications in prokaryotes. In this report, we demonstrate that cell-penetrating peptides (CPPs), both bacteria-expressed nona-arginine peptide (R9) and synthetic R9 (SR9), are able to deliver noncovalently associated proteins or quantum dots into four representative species of prokaryotes: cyanobacteria (Synechocystis sp. PCC 6803), bacteria (Escherichia coli DH5α and Arthrobacter ilicis D-50), and archaea (Thermus aquaticus). Although energy-dependent endocytosis is generally accepted as a hallmark that distinguishes eukaryotes from prokaryotes, cellular uptake of uncomplexed green fluorescent protein (GFP) by cyanobacteria was mediated by classical endocytosis. Mechanistic studies revealed that macropinocytosis plays a critical and major role in CPP-mediated protein transduction in all four prokaryotes. Membrane damage was not observed when cyanobacterial cells were treated with R9/GFP complexes, nor was cytotoxicity detected when bacteria or archaea were treated with SR9/QD complexes in the presence of macropinocytic inhibitors. These results indicate that the uptake of protein is not due to a compromise of membrane integrity in cyanobacteria, and that CPP can be an effective and safe carrier for membrane trafficking in prokaryotic cells. Our investigation provides important new insights into the transport of exogenous proteins and nanoparticles across the complex membrane systems of prokaryotes.

Department(s)

Biological Sciences

Keywords and Phrases

Archaeon; Bacterium; Cell Membrane; Endocytosis; Fluorescence Microscopy; Genetics; Metabolism; Permeability; Physiology; Protein Transport; Archaea; Bacteria; Cell-Penetrating Peptides (CPPs); Microscopy, Fluorescence; Prokaryotic Cells; Cyanobacteria; Macropinocytosis

International Standard Serial Number (ISSN)

0022-2631; 1432-1424

Document Type

Article - Journal

Document Version

Citation

File Type

text

Language(s)

English

Rights

© 2015 Springer Verlag, All rights reserved.

Publication Date

01 Apr 2015

PubMed ID

25655108

Link to Full Text

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