Interaction of dNTP, pyrophosphate and their analogs with the dNTP‐binding sites of E. coli DNA polymerase I Klenow fragment and human DNA polymerase

[摘要]

The 3',5'-exonuclease center of the Klenow fragment of E. coli DNA polymerase I (FK) was selectively blocked by NaF. The latter was shown to forbid the binding of nucleotides and their analogs to the enzyme exonuclease center. In the presence of poly(dT) · r(pA)₁₀ template · primer complex and NaF, we observed AMP, ADP, ATP, PP_i and dATP to be competitive inhibitors of the FK-catalyzed DNA polymerization. The interactions of the nucleotides with FK and human DNA polymerase α were compared to reveal similarity of binding to the DNA polymerizing centers. Structural components of dNTP and PP_i playing key roles in forming complexes with pro- and eukaryotic DNA polymerases were identified.

[学科分类] 生物化学/生物物理

[关键词] Klenow fragment;Human DNA polymerase α;Interaction with nucleotides;FK;Klenow fragment of E. coli DNA polymerase I;DPH;DNA polymerase α from human placenta