Monoclonal antibodies specific to human ETS-2 oncoprotein: recognition of epitopes clustered on the B domain.
Six monoclonal antibodies were prepared from mice immunized with a bacterially expressed human ets-2 protein. These antibodies specifically recognize the two human ets-2-encoded proteins p56 and p54 but failed to react with chicken, mouse, rat, bovine, or monkey proteins, suggesting that the antibodies recognize epitopes specific to the human ets-2 protein. Differential reactivities of these monoclonal antibodies with the peptide fragments generated by partial proteolytic digestion of the bacterially expressed ets-2 protein indicated that the six antibodies recognize at least three distinct epitopes in the B domain of the ets-2 protein. Immunoprecipitation experiments comparing native and denaturing conditions suggested that the ets-2 domain detected by the monoclonal antibodies is masked in the native condition by either protein folding or interacting proteins. The biochemical analysis of the ets-2 protein will be facilitated by the development of these monoclonal antibodies, which may be useful as both domain-specific probes and tools for specifically detecting the human ets-2 protein in heterologous expression systems.
Fujiwara
S
S
Laboratory of Molecular Oncology, National Cancer Institute, Frederick, MD.
Koizumi
S
S
Fisher
R J
RJ
Bhat
N K
NK
Papas
T S
TS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D000939:Epitopes; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007120:Immunochemistry; D051379:Mice; D010449:Peptide Mapping; D015682:Retroviridae Proteins, Oncogenic / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015682:Retroviridae Proteins, Oncogenic; C050379:oncogene proteins v-ets