Isolation of the catalytic core of DNA polymerase alpha from rabbit bone marrow

Nucleic Acids Res. 1982 Oct 11;10(19):6023-35. doi: 10.1093/nar/10.19.6023.

Abstract

Modification of the purification procedures for rabbit bone marrow DNA polymerase [Byrnes, J.J., & Black, V.L. (1978) Biochemistry 17, 4226-4231] has increased the yield and stability of the enzyme thus allowing further purification. In particular, the higher molecular weight form, alpha 1, has been more abundant. Additional purification has been obtained upon phosphocellulose and chromatofocusing column chromatography. SDS slab gel electrophoretic analyses of the eluates demonstrate a 135,000 molecular weight polypeptide in nearly pure form which correlates with DNA polymerase activity. Approximately 200,000 nmol of thymidine monophosphate is incorporated into DNA (mg of protein) -1h -1 at 37 degrees C. Similar to DNA polymerase alpha from other sources this enzyme is an acidic protein, is very sensitive to aphidicolin, and has no detectable 3' to 5' nuculease activity.

Publication types

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

MeSH terms

  • Animals
  • Aphidicolin
  • Bone Marrow / enzymology*
  • DNA Polymerase II / antagonists & inhibitors
  • DNA Polymerase II / isolation & purification*
  • DNA-Directed DNA Polymerase / isolation & purification*
  • Diterpenes / pharmacology
  • Kinetics
  • Molecular Weight
  • Rabbits

Substances

  • Diterpenes
  • Aphidicolin
  • DNA Polymerase II
  • DNA-Directed DNA Polymerase