title: 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.
authors: Department of Natural Resources and Environmental Studies, National Dong Hwa University, 1, Sec. 2, Da-Hsueh Road, Shoufeng Township, Hualien, 97401, Taiwan.
authors: Betty Revon
authors: BR
authors: Liu
authors: Yue-Wern
authors: YW
authors: Huang
authors: Robert S
authors: RS
authors: Aronstam
authors: Han-Jung
authors: HJ
authors: Lee
mesh_terms: D001105:Archaea / Q000378:metabolism; D001419:Bacteria / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D057846:Cell-Penetrating Peptides / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity; D004705:Endocytosis; D008856:Microscopy, Fluorescence; D010539:Permeability; D011387:Prokaryotic Cells / Q000502:physiology*; D021381:Protein Transport
chemical_list: D057846:Cell-Penetrating Peptides
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