Glycosylation of the hemagglutinin-neuraminidase glycoprotein of human parainfluenza virus type 1 affects its functional but not its antigenic properties.
The hemagglutinin-neuraminidase (HN) glycoprotein of human parainfluenza virus type 1 (hPIV-1) has been shown to be similar in predicted protein sequence and structure to those of Sendai virus, but it is more highly glycosylated. Because glycosylation can modify protein structure and function, we investigated the effect of glycosylation on the antigenic structure and biological function of the HN of hPIV-1. Antigenic and functional analyses were carried out with purified hPIV-1 virions treated with Endoglycosidase F, which removes carbohydrate moieties, because treatment of hPIV-1-infected LLC-MK2 cells with an inhibitor of glycosylation resulted in virions which were deficient in both HN and F surface glycoproteins. No change in the antigenic structure of the HN of hPIV-1 was detected after carbohydrate removal; epitope recognition by a panel of 7 hPIV-1 HN monoclonal antibodies (MAbs) was unchanged compared to untreated virions. Moreover, there was no change in the cross-reactivity of 8 of 10 Sendai virus HN MAbs, and only a slight change in the remaining 2. Nor did carbohydrate removal appear to affect hemagglutinating or neuraminidase activities; hemagglutination titers with chicken erythrocytes (cRBC) were unchanged, and in vitro neuraminidase activity with a small substrate (N-acetylneuraminlactose) showed only a 20% reduction. However, elution of deglycosylated hPIV-1 from agglutinated cRBC as a result of neuraminidase activity was reduced by 80%. These results suggest that the enzymatic activity of hPIV-1 HN was not directly affected by carbohydrate removal but that the reduction in elution was due to a change in the interaction of the HN with the host receptor. This was further supported by a 2- to 16-fold reduction in the ability of all 7 hPIV-1 HN MAbs to inhibit hemagglutination of deglycosylated hPIV-1 virus. Such a change in HN-host receptor interaction was found to involve a change in receptor specificity because deglycosylated virus was able to fully agglutinate cRBC stripped of receptors required by the native, glycosylated virus. We propose the following model for our results: deglycosylation of the HN of hPIV-1 causes the hemagglutinating portion of the molecule to recognize a new receptor which is not susceptible to enzymatic cleavage by the neuraminidase.
Gorman
W L
WL
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101.
Pridgen
C
C
Portner
A
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D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000737:chemistry / Q000502:physiology*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D015707:HN Protein / Q000378:metabolism*; D006801:Humans; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D010222:Parainfluenza Virus 1, Human / Q000187:drug effects / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D013329:Structure-Activity Relationship; D014415:Tunicamycin / Q000494:pharmacology; D014771:Virion / Q000187:drug effects / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D015707:HN Protein; D014415:Tunicamycin; D006026:Glycoside Hydrolases; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase