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Title: Chemical studies of viral entry mechanisms: I. Hydrophobic protein-lipid interactions during Sendai virus membrane fusion. II. Kinetics of bacteriophage. lambda. DNA injection

Abstract

Sendai virus glycoprotein interactions with target membranes during the early stages of fusion were examined using time-resolved hydrophobic photoaffinity labeling with the lipid-soluble carbene generator 3-(trifluoromethyl)-3-(m({sup 125}I) iodophenyl)diazirine. During Sendai virus fusion with liposomes composed of cardiolipin or phosphatidylserine, the viral fusion (F) protein is preferentially labeled at early time points, supporting the hypothesis that hydrophobic interaction of the fusion peptide at the N-terminus of the F{sub 1} subunit with the target membrane is an initiating event in fusion. Correlation of hydrophobic interactions with independently monitored fusion kinetics further supports this conclusion. The F{sub 1} subunit, containing the putative hydrophobic fusion sequence, is exclusively labeled, and the F{sub 2} subunit does not participate in fusion. Labeling shows temperature and pH dependence consistent with a need for protein conformational mobility and fusion at neutral pH. Higher amounts of labeling during fusion with CL vesicles than during virus-PS vesicle fusion reflects membrane packing regulation of peptide insertion into target membranes. Labeling of the viral hemagglutinin/neuraminidase (HN) at low pH indicates that HN-mediated fusion is triggered by hydrophobic interactions. Controls for diffusional labeling exclude a major contribution from this source. Labeling during reconstituted Sendai virus envelope-liposome fusion shows that functional reconstitution involves proteinmore » retention of the ability to undergo hydrophobic interactions. Examination of Sendai virus fusion with erythrocyte membranes indicates that hydrophobic interactions also trigger fusion between biological membranes. The data show that hydrophobic fusion protein interaction with both artificial and biological membranes is a triggering event in fusion.« less

Authors:
Publication Date:
Research Org.:
California Inst. of Tech., Pasadena, CA (USA)
OSTI Identifier:
6322404
Resource Type:
Miscellaneous
Resource Relation:
Other Information: Thesis (Ph. D.)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; GLYCOPROTEINS; BIOCHEMICAL REACTION KINETICS; VIRAL DISEASES; PATHOGENESIS; BACTERIOPHAGES; CELL MEMBRANES; DNA; ERYTHROCYTES; IODINE 125; LABELLING; TEMPERATURE DEPENDENCE; TRACER TECHNIQUES; VIRUSES; BETA DECAY RADIOISOTOPES; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY FLUIDS; CELL CONSTITUENTS; DAYS LIVING RADIOISOTOPES; DISEASES; ELECTRON CAPTURE RADIOISOTOPES; INFECTIOUS DISEASES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; KINETICS; MATERIALS; MEMBRANES; MICROORGANISMS; NUCLEI; NUCLEIC ACIDS; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; PARASITES; PROTEINS; RADIOISOTOPES; REACTION KINETICS; 550201* - Biochemistry- Tracer Techniques; 550901 - Pathology- Tracer Techniques

Citation Formats

Novick, S L. Chemical studies of viral entry mechanisms: I. Hydrophobic protein-lipid interactions during Sendai virus membrane fusion. II. Kinetics of bacteriophage. lambda. DNA injection. United States: N. p., 1990. Web.
Novick, S L. Chemical studies of viral entry mechanisms: I. Hydrophobic protein-lipid interactions during Sendai virus membrane fusion. II. Kinetics of bacteriophage. lambda. DNA injection. United States.
Novick, S L. 1990. "Chemical studies of viral entry mechanisms: I. Hydrophobic protein-lipid interactions during Sendai virus membrane fusion. II. Kinetics of bacteriophage. lambda. DNA injection". United States.
@article{osti_6322404,
title = {Chemical studies of viral entry mechanisms: I. Hydrophobic protein-lipid interactions during Sendai virus membrane fusion. II. Kinetics of bacteriophage. lambda. DNA injection},
author = {Novick, S L},
abstractNote = {Sendai virus glycoprotein interactions with target membranes during the early stages of fusion were examined using time-resolved hydrophobic photoaffinity labeling with the lipid-soluble carbene generator 3-(trifluoromethyl)-3-(m({sup 125}I) iodophenyl)diazirine. During Sendai virus fusion with liposomes composed of cardiolipin or phosphatidylserine, the viral fusion (F) protein is preferentially labeled at early time points, supporting the hypothesis that hydrophobic interaction of the fusion peptide at the N-terminus of the F{sub 1} subunit with the target membrane is an initiating event in fusion. Correlation of hydrophobic interactions with independently monitored fusion kinetics further supports this conclusion. The F{sub 1} subunit, containing the putative hydrophobic fusion sequence, is exclusively labeled, and the F{sub 2} subunit does not participate in fusion. Labeling shows temperature and pH dependence consistent with a need for protein conformational mobility and fusion at neutral pH. Higher amounts of labeling during fusion with CL vesicles than during virus-PS vesicle fusion reflects membrane packing regulation of peptide insertion into target membranes. Labeling of the viral hemagglutinin/neuraminidase (HN) at low pH indicates that HN-mediated fusion is triggered by hydrophobic interactions. Controls for diffusional labeling exclude a major contribution from this source. Labeling during reconstituted Sendai virus envelope-liposome fusion shows that functional reconstitution involves protein retention of the ability to undergo hydrophobic interactions. Examination of Sendai virus fusion with erythrocyte membranes indicates that hydrophobic interactions also trigger fusion between biological membranes. The data show that hydrophobic fusion protein interaction with both artificial and biological membranes is a triggering event in fusion.},
doi = {},
url = {https://www.osti.gov/biblio/6322404}, journal = {},
number = ,
volume = ,
place = {United States},
year = {Mon Jan 01 00:00:00 EST 1990},
month = {Mon Jan 01 00:00:00 EST 1990}
}

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