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The efficiency of interaction of deoxyribonucleosid‐5'‐mono‐, di‐ and triphosphates with the active centre of E. coli DNA polymerase I Klenow fragment
Author(s) -
Doronin S.V.,
Nevinsky G.A.,
Malygina T.O.,
Podust V.N.,
Khomov V.V.,
Lavrik O.I.
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)81500-5
Subject(s) - klenow fragment , nucleotide , dna polymerase i , chemistry , polymerase , dna polymerase , biochemistry , deoxyribonucleoside , enzyme , dna , stereochemistry , reverse transcriptase , exonuclease , polymerase chain reaction , gene
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.