Monoclonal antibody #5-2-26 recognizes the phosphatase-sensitive epitope of rabies virus nucleoprotein.
We prepared monoclonal antibodies (MAbs) against the rabies virus N protein, among which one antibody (MAb 5-2-26) was shown to lack reactivity with the phosphatase-treated N protein. The MAb was able to recognize the sodium dodecyl sulfate (SDS)-denatured N protein. The MAb did not recognize the N-protein analogues produced in Escherichia coli (E. coli), indicating that the N-gene products were not normally processed in E. coli after translation. On the other hand, the MAb reacted normally with N-gene products produced in COS-7 cells, but not with those produced in the presence of K-252a (a protein kinase inhibitor of a broad spectrum). The MAb displayed weak cross-reactivity with the Triton-insoluble network structures composed of several components, while another phosphoprotein (M1) of the virus was not recognized at all. These results suggest that MAb 5-2-26 preferentially recognizes a phosphatase-sensitive linear epitope of N protein, which may enable further investigations to be conducted on the mechanism of N-protein phosphorylation and its role(s) in virus replication.
Kawai
A
A
Department of Molecular Microbiology, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Anzai
J
J
Honda
Y
Y
Morimoto
K
K
Takeuchi
K
K
Kohno
T
T
Wakisaka
K
K
Goto
H
H
Minamoto
N
N
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology; D019556:COS Cells; D002227:Carbazoles / Q000494:pharmacology; D002460:Cell Line; D006224:Cricetinae; D003429:Cross Reactions; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000939:Epitopes / Q000276:immunology* / Q000378:metabolism; D026121:Indole Alkaloids; D051379:Mice; D008807:Mice, Inbred BALB C; D019251:Nucleocapsid / Q000276:immunology*; D019590:Nucleocapsid Proteins; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011820:Rabies virus / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D002227:Carbazoles; D004791:Enzyme Inhibitors; D000939:Epitopes; D026121:Indole Alkaloids; D019590:Nucleocapsid Proteins; C099601:nucleocapsid protein, Rabies virus; C049985:staurosporine aglycone; D011493:Protein Kinase C; D010744:Phosphoric Monoester Hydrolases