The efficiency of interaction of deoxyribonucleosid‐5'‐mono‐, di‐ and triphosphates with the active centre of E. coli DNA polymerase I Klenow fragment

FEBS Letters

Lavrik, O.I.²  Doronin, S.V.²  Malygina, T.O.²  Podust, V.N.²  Khomov, V.V.¹  Nevinsky, G.A.² 

[1] Scientific Research Institute of Design and Technology of Biologically Active Substances, Berdsk, Novosibirsk Region 633190, USSR;Institute of Bioorganic Chemistry, Siberian Division of the USSR Academy of Sciences, Novosibirsk 630090, USSR

关键词: DNA polymerase I Klenow fragment;    dNTP selection;    Affinity inhibition;    (Escherichia coli);    epATP;    adenosine 2';    3 '-riboepoxide 5 '-triphosphate;    BSA;    bovine serum albumin;    FK;    E. coli DNA polymerase I Klenow fragment;    DPH;    DNA polymerase α from human placenta;   

DOI  :  10.1016/0014-5793(89)81500-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The interaction of deoxyribonucleoside-5'-mono-, di- and triphosphates with E. coli DNA polymerase I Klenow fragments was examined. Dissociation constants of the enzyme complex with nucleotides were determined from the data on the enzyme inactivation by adenosine 2',3'-riboepoxide 5'-triphosphate. The role of nucleotide bases, phosphate groups and sugar moieties in the complex formation of nucleotides with the enzyme was elucidated. The necessity of complementary interaction of nucleotides with templates for template-controlled ‘adjusting’ of complementary dNTP to its reactive state was found. The crucial role of the interaction of dNTP γ-phosphate with the enzyme in this process is discussed.

【 授权许可】

Unknown