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  • Nonviral Transfer Technology
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Novel cell permeable motif derived from the PreS2-domain of hepatitis-B virus surface antigens

Gene Therapy volume 7, pages 750–758 (2000)Cite this article

Abstract

Efficient transfer of proteins or nucleic acids across cellular membranes is a major problem in cell biology. Recently the existence of a fusogenic sequence was predicted in the junction area of the PreS2- and S-domain of the hepatitis-B virus surface antigens. We have identified cell permeability as a novel property of the PreS2-domain. Cell permeability of PreS2 is not restricted to hepatocytes. PreS2 translocates in an energy-independent manner into cells and is evenly distributed over the cytosol. Detailed analysis revealed that cell-permeability is mediated by an amphipatic α-helix between amino acids 41 and 52 of PreS2. Destruction of this translocation motif (PreS2-TLM) abolishes cell permeability. PreS2-TLM per se can act as a shuttle for peptides and functional proteins (such as EGFP). This permits the highly specific modulation of intracellular signal transduction by transfer of peptides competing protein–protein interactions as demonstrated by specific inhibition of TNFα-dependent activation of c-Raf-1 kinase. Moreover in vivo functionality was demonstrated by PreS2-TLM-dependent protein transfer into primary bone marrow cells and into the liver. The amphipatic motif is conserved between the different hepatitis-B virus subtypes, and the surface proteins of avian and rodent hepadnaviruses exhibit similar amphipatic peptide sequences. In respect to hepatitis-B virus-infection, the PreS2-TLM could represent the postulated fusion peptide and play a crucial role in the internalization of the viral particle.

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Acknowledgements

Antibodies were kindly provided by Dr L Mimms, Chicago (HBV 25–19), Professor W Gerlich, Gieβen (Q19–10) and Dr A Budkowska, Paris (F124). We thank Dr C Plank, Institute for Experimental Oncology, Munich, for peptide synthesis, Professor M Wink, Department for Pharmaceutical Biology, Heidelberg, for callus cultures of lupinus albus and his kind invitation to carry out the experiments in his laboratory and Dr B Munz, Stanford, for helpful discussion. We are indebted to Professor PH Hofschneider, Martinsried, for his continuous generous support. This work was supported by the Deutsche Stiftung für Krebsforschung and the MMW (Munich).

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  1. Max-Planck-Institut für Biochemie, AG Virusforschung, Martinsried, Germany

    S Oess

  2. Institut für Experimentelle Onkologie und Therapieforschung der Technischen Universität München, Germany

    S Oess & E Hildt

  3. Robert Koch-Institut, Berlin, Germany

    E Hildt

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Oess, S., Hildt, E. Novel cell permeable motif derived from the PreS2-domain of hepatitis-B virus surface antigens. Gene Ther 7, 750–758 (2000). https://doi.org/10.1038/sj.gt.3301154

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