Premium
DNA polymerase I (Klenow fragment): Role of the structure and length of a template in enzyme recognition
Author(s) -
Kolocheva T.I.,
Nevinsky G.A.,
Volchkova V.A.,
Levina A.S.,
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)80439-9
Subject(s) - klenow fragment , dna polymerase i , oligonucleotide , chemistry , dna , stereochemistry , dna polymerase , polymerase , polynucleotide , nucleotide , enzyme , biochemistry , microbiology and biotechnology , biology , polymerase chain reaction , reverse transcriptase , exonuclease , gene
The values of K d and Gibbs energy (ΔG°) 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 , d(pG) 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) 2 C[Pt 2+ (NH 3 ) 2 OH](pT) 7 and d[(Tp) 2 C(Pt 2+ (NH 3 ) 2 OH)p] 3 T 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 Me 2+ ‐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.