Dodging cellular customs: Smuggling macromolecules into hepatocytes. Bagai S, Sarkar DP. Fusion-mediated microinjection of lysozyme into HepG2 cells through hemagglutinin neuraminidase-depleted Sendai virus envelopes. J Biol Chem 1994;269:1966-1972

Soumit K. Basu, Jayanta Roy-Chowdhury

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The potential of reconstitued Sendai viral envelopes containing only the fusion protein (F-virosomes) was evaluated for a targeted cytosolic delivery of lysozyme to human hepatoblastoma cells (HepG2) in culture. 125I-Lysozyme loaded into F-virosomes was used to monitor its fusion-mediated transfer to the HepG2 cells. Using fusion assay based on the transfer of water soluble probe, we have demonstrated the existence of aqueous connection between F-virosomes and target cells. Target specificity of the F-virosomes was ensured by the strong interaction between terminal β-galactose moiety of F protein and the asialogly-coprotein receptor on the membrane of HepG2 cells. Incubation of the loaded F-virosomes with cells resulted in fusion-mediated injection, as inferred from the ability of cells to internalize lysozyme in the presence of azide (an inhibitor of the endocytotic process). Binding as well as fusion of the F-virosomes to HepG2 cells was solely mediated by the F protein. Introduction of 125I-lysozyme into the HepG2 cells was confirmed by selective accumulation of acid and antibody-precipitable radioactivity in the cytosolic compartment. The structural integrity of the internalized lysozyme was also assessed. The potential usefulness of F-virosomes with defined specificities as biological carrierfor both in vitro and in vivo cytosolic delivery of macromolecules and drugs has been established.

Original languageEnglish (US)
Pages (from-to)1640-1642
Number of pages3
JournalHepatology
Volume20
Issue number6
StatePublished - Dec 1994

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Virosomes
Sendai virus
Hep G2 Cells
Hemagglutinins
Neuraminidase
Microinjections
Muramidase
Hepatocytes
Hepatoblastoma
Proteins
Azides
Galactose
Radioactivity
Injections
Acids
Membranes
Water
Antibodies

ASJC Scopus subject areas

  • Hepatology

Cite this

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title = "Dodging cellular customs: Smuggling macromolecules into hepatocytes. Bagai S, Sarkar DP. Fusion-mediated microinjection of lysozyme into HepG2 cells through hemagglutinin neuraminidase-depleted Sendai virus envelopes. J Biol Chem 1994;269:1966-1972",
abstract = "The potential of reconstitued Sendai viral envelopes containing only the fusion protein (F-virosomes) was evaluated for a targeted cytosolic delivery of lysozyme to human hepatoblastoma cells (HepG2) in culture. 125I-Lysozyme loaded into F-virosomes was used to monitor its fusion-mediated transfer to the HepG2 cells. Using fusion assay based on the transfer of water soluble probe, we have demonstrated the existence of aqueous connection between F-virosomes and target cells. Target specificity of the F-virosomes was ensured by the strong interaction between terminal β-galactose moiety of F protein and the asialogly-coprotein receptor on the membrane of HepG2 cells. Incubation of the loaded F-virosomes with cells resulted in fusion-mediated injection, as inferred from the ability of cells to internalize lysozyme in the presence of azide (an inhibitor of the endocytotic process). Binding as well as fusion of the F-virosomes to HepG2 cells was solely mediated by the F protein. Introduction of 125I-lysozyme into the HepG2 cells was confirmed by selective accumulation of acid and antibody-precipitable radioactivity in the cytosolic compartment. The structural integrity of the internalized lysozyme was also assessed. The potential usefulness of F-virosomes with defined specificities as biological carrierfor both in vitro and in vivo cytosolic delivery of macromolecules and drugs has been established.",
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