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Physicochemical properties of pseudorabies virus hemagglutinin
1Department of Veterinary Epizootiology, College of Agriculture and Veterinary Medicine, Nihon Univesity, Kanagawa, Japan.
Abstract:
Pseudorabies virus hemagglutinin was readily adsorbed on mouse erythrocytes at 4, 22, or 37 degrees C, but not on cattle erythrocytes. The adsorbed hemagglutinin could not be eluted from the cells by resuspending in phosphate-buffered saline (PBS), by incubating at 37 or 50 degrees C, or by incubating in the presence of neuraminidase. The receptor on mouse erythrocytes for the hemagglutinin was inactivated by trypsin, but not by neuraminidase, sodium deoxycholate (DOC), potassium periodate (KIO4), dithiothreitol (DTT), 2-mercaptoethanol (2-ME) and formalin. The hemagglutinin was inactivated by trypsin, alpha-amylase, pepsin, DOC, KIO4, and ethylendiamine-tetraacetic acid (EDTA), but not by papain, beta-glucosidase, phospholipase C, neuraminidase, DTT, 2-ME, Tween-80, ethylether, chloroform, trichloro-trifluoroethane, beta-propiolactone and formalin, suggesting that the hemagglutinin active component involved glycoproteins. The hemagglutinin was stable at 37 degrees C for lower temperatures but not at 60 degrees C or higher. The hemagglutinin activity was resistant to ultraviolet irradiation, while the infectivity was very susceptible. The hemagglutinin and the infectivity were readily sedimented by ultracentrifugation at 48,000 x g for 3 hr. In rate zonal centrifugation of the preparation on a sucrose density gradient, the hemagglutination (HA) activity showed a sharp peak at 1.22 g/ml coinciding with the peak of infectivity. The HA activity in the peak fraction seemed to be structually associated with virus particles. After fractionation of the virus by Nonidet P-40, the HA activity was found only in the fraction of the envelope material, indicating that the hemagglutinin is situated in the viral envelop.
Insights
Pseudorabies virus hemagglutinin binds mouse, but not cattle, red blood cells. This hemagglutinin, likely a glycoprotein, is located in the viral envelope and is sensitive to heat and certain enzymes.
Area of Science:
- Virology
- Molecular Biology
- Immunology
Background:
- Pseudorabies virus (PRV) is an important pathogen affecting swine.
- Hemagglutinin (HA) is a key viral surface protein involved in host cell attachment.
Purpose of the Study:
- To characterize the properties of PRV hemagglutinin.
- To determine the cellular receptor and the nature of the hemagglutinin molecule.
Main Methods:
- Erythrocyte adsorption assays at various temperatures.
- Elution and inactivation studies using different chemical and enzymatic treatments.
- Ultracentrifugation and sucrose density gradient centrifugation.
- Virus fractionation using Nonidet P-40.
Main Results:
- PRV hemagglutinin adsorbed to mouse erythrocytes but not cattle erythrocytes.
- The receptor on mouse erythrocytes was inactivated by trypsin, suggesting a proteinaceous nature.
- The hemagglutinin itself was inactivated by proteases, alpha-amylase, and detergents, indicating a glycoprotein structure.
- Hemagglutinin was heat-labile above 60°C and resistant to UV irradiation.
- Hemagglutination activity was associated with virus particles and located in the viral envelope fraction.
Conclusions:
- PRV hemagglutinin exhibits specific binding to mouse erythrocytes.
- The viral hemagglutinin is a glycoprotein component of the PRV envelope.
- These findings contribute to understanding PRV-host interactions and viral structure.