Mutation of arginine residues to avoid non-specific cellular uptakes for hepatitis B virus core particles.

Kenta Morita,Jun Ishii,Khuloud T Al-Jamal,Yuya Nishimura,Izzat Fahimuddin Bin Mohamed Suffian,Sachiko Nakamura-Tsuruta,Chiaki Ogino,Akihiko Kondo

doi:10.1186/s12951-015-0074-8

Kenta Morita, Jun Ishii + Show 6 more

Open Access

https://doi.org/10.1186/s12951-015-0074-8

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Abstract

BackgroundThe hepatitis B virus core (HBc) particle is known as a promising new carrier for the delivery of drugs and nucleic acids. However, since the arginine-rich domain that is located in the C-terminal region of the HBc monomer binds to the heparan sulphate proteoglycan on the cell surface due to its positive charge, HBc particles are introduced non-specifically into a wide range of cells. To avoid non-specific cellular uptake with the intent to control the ability of cell targeting, we individually replaced the respective arginine (R) residues of the arginine-rich domain located in amino acid positions 150–159 in glycine (G) residues.ResultsThe mutated HBc particles in which R154 was replaced with glycine (G) residue (R154G) showed a drastic decrease in the ability to bind to the heparan sulphate proteoglycan and to avoid non-specific cellular uptake by several types of cancer cells.ConclusionsBecause this mutant particle retains most of its C-terminal arginine-rich residues, it would be useful in the targeting of specificity-altered HBc particles in the delivery of nucleic acids.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-015-0074-8) contains supplementary material, which is available to authorized users.

Highlights

The hepatitis B virus core (HBc) particle is known as a promising new carrier for the delivery of drugs and nucleic acids
HBc monomers are composed of two distinct domains: i) an assembly domain (amino acid residues 1–149) that drives particle formation, and ii) an arginine-rich domain that recognizes the cell surface heparan sulphate proteoglycan with an electrostatic interaction [6]
The particle formation was confirmed by atomic force microscopy (AFM), scanning electron microscope (SEM) and dynamic light scattering (DLS) (Figure 1)

Summary

Introduction

The hepatitis B virus core (HBc) particle is known as a promising new carrier for the delivery of drugs and nucleic acids. Since the arginine-rich domain that is located in the C-terminal region of the HBc monomer binds to the heparan sulphate proteoglycan on the cell surface due to its positive charge, HBc particles are introduced non- into a wide range of cells. To avoid non-specific cellular uptake with the intent to control the ability of cell targeting, we individually replaced the respective arginine (R) residues of the arginine-rich domain located in amino acid positions 150–159 in glycine (G) residues. HBc monomers are composed of two distinct domains: i) an assembly domain (amino acid residues (aa) 1–149) that drives particle formation, and ii) an arginine-rich domain (aa 150–183) that recognizes the cell surface heparan sulphate proteoglycan with an electrostatic interaction [6]. The argininerich domain behaves as a binding site for nucleic acids, because of its positively charged residues [8,9]

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Conclusion