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Crystal structures of the Klenow fragment of Thermus aquaticus DNA polymerase I complexed with deoxyribonucleoside triphosphates
Author(s) -
Li Ying,
Kong Yong,
Korolev Sergey,
Waksman Gabriel
Publication year - 1998
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560070505
Subject(s) - klenow fragment , thermus aquaticus , deoxyribonucleoside , taq polymerase , polymerase , dna polymerase , chemistry , base pair , thermus thermophilus , dna polymerase i , stereochemistry , dna , nucleotide , primer (cosmetics) , microbiology and biotechnology , biochemistry , biology , exonuclease , polymerase chain reaction , reverse transcriptase , escherichia coli , organic chemistry , gene
The crystal structures of the Klenow fragment of the Thermus aquaticus DNA polymerase I (Klentaq 1) complexed with four deoxyribonucleoside triphosphates (dNTP) have been determined to 2.5 Å resolution. The dNTPs bind adjacent to the O helix of Klentaq 1. The triphosphate moieties are at nearly identical positions in all four complexes and are anchored by three positively charged residues, Arg659, Lys663, and Arg587, and by two polar residues, His639 and Gln613. The configuration of the base moieties in the Klentaq 1/dNTP complexes demonstrates variability suggesting that dNTP binding is primarily determined by recognition and binding of the phosphate moiety. However, when superimposed on the Taq polymerase/blunt end DNA complex structure (Eom et al., 1996), two of the dNTP/Klentaql structures demonstrate appropriate stacking of the nucleotide base with the 3′ end of the DNA primer strand, suggesting that at least in these two binary complexes, the observed dNTP conformations are functionally relevant.