Probing DNA polymerase alpha with monoclonal antibodies.
DNA polymerase alpha was purified from human KB cells by immunoaffinity chromatography. Enzyme activity was inhibited by three different monoclonal antibodies (SJK-132, SJK-211, SJK-287). Kinetic analysis showed that each antibody neutralized polymerase activity by a different mechanism. SJK-132 was competitive with DNA indicating it interacts with the DNA binding domain of the polymerase. SJK-287 showed a biphasic response to dCTP suggesting two dCTP binding sites exist on polymerase alpha. SJK-211 was non-competitive with DNA, dCTP and dATP.
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D004257:DNA Polymerase II / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007624:KB Cells; D007700:Kinetics
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D004257:DNA Polymerase II