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Title: Radiation inactivation analysis of influenza virus reveals different target sizes for fusion, leakage, and neuraminidase activities

Abstract

The size of the functional units responsible for several activities carried out by the influenza virus envelope glycoproteins was determined by radiation inactivation analysis. Neuraminidase activity, which resides in the glycoprotein NA, was inactivated exponentially with an increasing radiation dose, yielding a target size of 94 +/- 5 kilodaltons (kDa), in reasonable agreement with that of the disulfide-bonded dimer (120 kDa). All the other activities studied are properties of the HA glycoprotein and were normalized to the known molecular weight of the neuraminidase dimer. Virus-induced fusion activity was measured by two phospholipid dilution assays: relief of energy transfer between N-(7-nitro-2,1,3-benzoxadiazol-4-yl)dipalmitoyl-L-alpha- phosphatidylethanolamine (N-NBD-PE) and N-(lissamine rhodamine B sulfonyl)-dioleoyl-L-alpha-phosphatidylethanolamine (N-Rh-PE) in target liposomes and relief of self-quenching of N-Rh-PE in target liposomes. Radiation inactivation of fusion activity proceeded exponentially with radiation dose, yielding normalized target sizes of 68 +/- 6 kDa by assay i and 70 +/- 4 kDa by assay ii. These values are close to the molecular weight of a single disulfide-bonded (HA1 + HA2) unit (75 kDa), the monomer of the HA trimer. A single monomer is thus inactivated by each radiation event, and each monomer (or some part of it) constitutes a minimal functional unit capable of mediatingmore » fusion. Virus-induced leakage of calcein from target liposomes and virus-induced leakage of hemoglobin from erythrocytes (hemolysis) both showed more complex inactivation behavior: a pronounced shoulder was present in both inactivation curves, followed by a steep drop in activity at higher radiation levels.« less

Authors:
; ; ;
Publication Date:
Research Org.:
Univ. of Medicine and Dentistry of New Jersey, Piscataway
OSTI Identifier:
6850477
Alternate Identifier(s):
OSTI ID: 6850477
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemistry; (United States); Journal Volume: 20
Country of Publication:
United States
Language:
English
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; GLUCOPROTEINS; BIOLOGICAL FUNCTIONS; O-GLYCOSYL HYDROLASES; ENZYME ACTIVITY; VIRUSES; BIOLOGICAL RADIATION EFFECTS; INACTIVATION; BIOCHEMISTRY; DOSE-RESPONSE RELATIONSHIPS; ENERGY TRANSFER; HEMOLYSIS; IONIZING RADIATIONS; LIPOSOMES; MOLECULAR WEIGHT; RADIATION DOSES; BIOLOGICAL EFFECTS; CARBOHYDRATES; CELL CONSTITUENTS; CHEMISTRY; DOSES; ENZYMES; FUNCTIONS; GLYCOSYL HYDROLASES; HYDROLASES; LYSIS; MICROORGANISMS; ORGANIC COMPOUNDS; ORGANOIDS; PARASITES; PATHOLOGICAL CHANGES; PROTEINS; RADIATION EFFECTS; RADIATIONS; SACCHARIDES 560130* -- Radiation Effects on Microorganisms

Citation Formats

Gibson, S., Jung, C.Y., Takahashi, M., and Lenard, J. Radiation inactivation analysis of influenza virus reveals different target sizes for fusion, leakage, and neuraminidase activities. United States: N. p., 1986. Web. doi:10.1021/bi00368a064.
Gibson, S., Jung, C.Y., Takahashi, M., & Lenard, J. Radiation inactivation analysis of influenza virus reveals different target sizes for fusion, leakage, and neuraminidase activities. United States. doi:10.1021/bi00368a064.
Gibson, S., Jung, C.Y., Takahashi, M., and Lenard, J. Tue . "Radiation inactivation analysis of influenza virus reveals different target sizes for fusion, leakage, and neuraminidase activities". United States. doi:10.1021/bi00368a064.
@article{osti_6850477,
title = {Radiation inactivation analysis of influenza virus reveals different target sizes for fusion, leakage, and neuraminidase activities},
author = {Gibson, S. and Jung, C.Y. and Takahashi, M. and Lenard, J.},
abstractNote = {The size of the functional units responsible for several activities carried out by the influenza virus envelope glycoproteins was determined by radiation inactivation analysis. Neuraminidase activity, which resides in the glycoprotein NA, was inactivated exponentially with an increasing radiation dose, yielding a target size of 94 +/- 5 kilodaltons (kDa), in reasonable agreement with that of the disulfide-bonded dimer (120 kDa). All the other activities studied are properties of the HA glycoprotein and were normalized to the known molecular weight of the neuraminidase dimer. Virus-induced fusion activity was measured by two phospholipid dilution assays: relief of energy transfer between N-(7-nitro-2,1,3-benzoxadiazol-4-yl)dipalmitoyl-L-alpha- phosphatidylethanolamine (N-NBD-PE) and N-(lissamine rhodamine B sulfonyl)-dioleoyl-L-alpha-phosphatidylethanolamine (N-Rh-PE) in target liposomes and relief of self-quenching of N-Rh-PE in target liposomes. Radiation inactivation of fusion activity proceeded exponentially with radiation dose, yielding normalized target sizes of 68 +/- 6 kDa by assay i and 70 +/- 4 kDa by assay ii. These values are close to the molecular weight of a single disulfide-bonded (HA1 + HA2) unit (75 kDa), the monomer of the HA trimer. A single monomer is thus inactivated by each radiation event, and each monomer (or some part of it) constitutes a minimal functional unit capable of mediating fusion. Virus-induced leakage of calcein from target liposomes and virus-induced leakage of hemoglobin from erythrocytes (hemolysis) both showed more complex inactivation behavior: a pronounced shoulder was present in both inactivation curves, followed by a steep drop in activity at higher radiation levels.},
doi = {10.1021/bi00368a064},
journal = {Biochemistry; (United States)},
number = ,
volume = 20,
place = {United States},
year = {Tue Oct 07 00:00:00 EDT 1986},
month = {Tue Oct 07 00:00:00 EDT 1986}
}