DNA polymerase I (Klenow fragment): role of the structure and length of a template in enzyme recognition

FEBS Lett. 1989 May 8;248(1-2):97-100. doi: 10.1016/0014-5793(89)80439-9.


The values of Kd and Gibbs energy (delta G degrees) have been measured for complexes of the template site of DNA polymerase I Klenow fragment with the homo-oligonucleotides d(pC)n, d(pT)n, and d(pA)n and hetero-oligonucleotides of various structures and lengths. These parameters were evaluated from the protective effect of the oligonucleotide on enzyme inactivation by the affinity reagents d(Tp)2C[Pt2+ (NH3)2OH](pT)7 and d[(Tp2)C(Pt2+(NH3)2OH)p]3T of the template site. The present results and previously reported data [(1985) Biorg. Khim. 13, 357-369] indicate that the nucleoside components of the template form complexes as a result of their hydrophobic interactions with the enzyme. Only one template internucleotide phosphate forms an Me2+-dependent electrostatic contact and a hydrogen bond with the enzyme. The 19-20-nucleotide fragments of the template appear to interact with the protein molecule.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Polymerase I*
  • DNA*
  • DNA-Directed DNA Polymerase*
  • Electrochemistry
  • Hydrogen Bonding
  • Kinetics
  • Models, Theoretical
  • Molecular Structure
  • Oligonucleotides
  • Peptide Fragments
  • Templates, Genetic


  • Oligonucleotides
  • Peptide Fragments
  • DNA
  • DNA Polymerase I
  • DNA-Directed DNA Polymerase