| Abstract: | recA protein of Escherichia coli promotes a wide variety of DNA reactions in vitro. Specific effectors of recA protein should be very useful in elucidating the mechanisms of these complex reactions. Six mouse hybridoma clones that secreted class G immunoglobulins specific to recA protein were obtained in three cell-fusion experiments. Five IgGs were purified by affinity chromatography. These monoclonal antibodies were characterized by examining their effects on the single-stranded DNA-dependent ATPase activity, negatively superhelical double-stranded DNA-dependent ATPase activity, and an activity in pairing negatively superhelical closed circular double-stranded DNA and homologous single-stranded DNA-fragments to form D-loops. These IgGs inhibited all, some, or one of these three activities, and from the spectra of their inhibitory effects they were classified into four groups. This classification suggests that each of the monoclonal antibodies binds to one of at least four antigenic determinants on recA protein and specifically inactivates one or more of the active centers on the protein. These monoclonal antibodies will be useful in analyzing the complex reactions promoted by recA protein. |
