Document Detail


Fusion-mediated microinjection of lysozyme into HepG2 cells through hemagglutinin neuraminidase-depleted Sendai virus envelopes.
MedLine Citation:
PMID:  8294448     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The potential of reconstituted 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 beta-galactose moiety of F protein and the asialoglycoprotein 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 carrier for both in vitro and in vivo cytosolic delivery of macromolecules and drugs has been established.
Authors:
S Bagai; D P Sarkar
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  269     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1994 Jan 
Date Detail:
Created Date:  1994-02-25     Completed Date:  1994-02-25     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1966-72     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, University of Delhi South Campus, New Delhi, India.
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MeSH Terms
Descriptor/Qualifier:
Asialoglycoprotein Receptor
Asialoglycoproteins / metabolism
Biological Transport
Cell Line
Galactose
Hepatoblastoma
Humans
Kinetics
Liver Neoplasms
Membrane Fusion
Microinjections
Muramidase / administration & dosage*,  metabolism*
Neuraminidase
Parainfluenza Virus 1, Human
Receptors, Cell Surface / metabolism*
Subcellular Fractions / metabolism
Tumor Cells, Cultured
Viral Fusion Proteins*
Chemical
Reg. No./Substance:
0/Asialoglycoprotein Receptor; 0/Asialoglycoproteins; 0/Receptors, Cell Surface; 0/Viral Fusion Proteins; 26566-61-0/Galactose; EC 3.2.1.17/Muramidase; EC 3.2.1.18/Neuraminidase

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