A cell fusion-inhibiting monoclonal antibody binds to the presumed stalk domain of the human parainfluenza type 2 virus hemagglutinin-neuraminidase protein.
Previously, we obtained a neutralizing monoclonal antibody directed against the hemagglutinin-neuraminidase (HN) protein of human parainfluenza type 2 virus (PIV2), which was able to prevent cell fusion without affecting the hemagglutinating and neuraminidase activities. In this study, four escape mutants of PIV2 have been obtained under pressure of the monoclonal antibody. Intriguingly, the HN protein of each mutant proved to have two amino acid substitutions, one of which is at 83Asn or 91Lys, and another one is at 150Leu, 160Ala, or 186Met. One mutant designated F13, which has substitutions at 83Asn and 186Met in the HN protein, could not cause cell fusion in HeLa cells despite its multiple replication, while the other mutants formed typical syncytial cells. The deduced amino acid sequence of F13 fusion (F) protein proved to be identical to that of wild-type F protein, and furthermore, protein expression analyses have revealed that the low-fusion phenotype of F13 was due to its mutated HN protein, whose antigenicity to the monoclonal antibody was abolished by the single mutation at 83Asn. These observations have suggested that the principal epitope for the monoclonal antibody resides in the presumed stalk domain of the HN protein, which may play an important role in promoting cell fusion.
Yuasa
T
T
Department of Microbiology, Mie University School of Medicine, Japan.
Kawano
M
M
Tabata
N
N
Nishio
M
M
Kusagawa
S
S
Komada
H
H
Matsumura
H
H
Ito
Y
Y
Tsurudome
M
M
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D001665:Binding Sites; D002459:Cell Fusion / Q000187:drug effects / Q000502:physiology*; D003062:Codon; D000939:Epitopes / Q000378:metabolism; D015726:Giant Cells; D015707:HN Protein / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D016297:Mutagenesis, Site-Directed; D010223:Parainfluenza Virus 2, Human / Q000378:metabolism / Q000502:physiology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D003062:Codon; D000939:Epitopes; D015707:HN Protein; D011994:Recombinant Proteins