| Cellular cytoplasmic delivery of a polypeptide toxin by reconstituted influenza virus envelopes (virosomes). | |
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MedLine Citation:
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PMID: 8049214 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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In this paper it is demonstrated that reconstituted influenza virus envelopes (virosomes) fuse efficiently with membranes of the endosomal cell compartment of cultured cells, after internalization through receptor-mediated endocytosis. As a consequence, molecules, encapsulated in the virosomal interior, are transferred to the cell cytoplasm. This process was monitored on the basis of delivery of subunit A of diphtheria toxin (DTA), initially encapsulated in the virosomal lumen. Virosome-mediated cytoplasmic delivery of DTA resulted in a virtually complete inhibition of cellular protein synthesis. DTA delivery was blocked by factors inhibiting the pH-dependent fusion activity of viral hemagglutinin, i.e., 20 mM NH4Cl, preincubation of the virosomes at low pH, and anti-hemagglutinin antibodies. Quantitation of the extent of virosome-mediated delivery of biologically active DTA demonstrated that a lower bound of approximately 10% of the virosomes entering the cells deposited their aqueous contents into the cytosol. This is in good agreement with the final extent of virosome fusion of 40%, as determined by a fluorescence lipid mixing assay based on the dilution of pyrene-labeled phosphatidylcholine from the virosomal into the endosomal membrane. At an added concentration of 1 microM virosomal lipid per 50,000 cells, depending on the condition of the experiment, a minimum of 4000 virosomes per cell were found to be internalized. Virosomes could also be induced to fuse with the cell plasma membrane by a transient lowering of the extracellular pH, as detected by appearance of virosome-encapsulated DTA in the cytoplasm.(ABSTRACT TRUNCATED AT 250 WORDS) |
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Authors:
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R Bron; A Ortiz; J Wilschut |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Biochemistry Volume: 33 ISSN: 0006-2960 ISO Abbreviation: Biochemistry Publication Date: 1994 Aug |
Date Detail:
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Created Date: 1994-09-08 Completed Date: 1994-09-08 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0370623 Medline TA: Biochemistry Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 9110-7 Citation Subset: IM |
Affiliation:
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Department of Physiological Chemistry, University of Groningen, The Netherlands. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Line Cell Membrane / metabolism Chick Embryo Cricetinae Cytoplasm / metabolism Diphtheria Toxin / metabolism*, toxicity Drug Carriers Influenza A virus / physiology* Kidney Kinetics Phosphatidylcholines Protein Synthesis Inhibitors / metabolism, toxicity Viral Envelope Proteins / biosynthesis, metabolism* |
| Chemical | |
Reg. No./Substance:
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0/Diphtheria Toxin; 0/Drug Carriers; 0/Phosphatidylcholines; 0/Protein Synthesis Inhibitors; 0/Viral Envelope Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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