Probing DNA polymerase alpha with monoclonal antibodies

FEBS Lett. 1987 Feb 23;212(2):259-62. doi: 10.1016/0014-5793(87)81356-x.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies, Monoclonal
  • Antigen-Antibody Complex
  • DNA Polymerase II / immunology
  • DNA Polymerase II / metabolism*
  • Humans
  • KB Cells
  • Kinetics

Substances

  • Antibodies, Monoclonal
  • Antigen-Antibody Complex
  • DNA Polymerase II