title: Thermostable DNA polymerase from Thermus thermophilus B35: influence of divalent metal ions on the interaction with deoxynucleoside triphosphates.
abstract: The interaction of DNA polymerase from Thermus thermophilus B35 (Tte-pol) with deoxynucleoside triphosphates in the presence of different divalent metal ions has been studied. DNA synthesis and competitive inhibition of the polymerase reaction by non-complementary dNTPs are described with corresponding kinetic schemes. The co-factor properties of some metals (Mg2+, Mn2+, Co2+, Ni2+, Cu2+, Ca2+, Cd2+, and Zn2+) were investigated, and their activating concentration ranges were determined. It was found that kcat values are significantly decreased and Km values slowly decrease when Mn2+ displaces Mg2+. The value of Kd for DNA template-primer is Me2+-independent, whereas Kd values for non-complementary dNTPs decrease in the presence of Mn2+. Tte-pol processivity but not DNA synthesis efficiency is Me2+-type independent.
authors.affiliation: Novosibirsk Institute of Bioorganic Chemistry, Siberian Branch of the Russian Academy of Sciences, Novosibirsk. nadyarec@niboch.nsc.ru
authors.forename: N I
authors.initials: NI
authors.lastname: Rechkunova
authors.forename: S S
authors.initials: SS
authors.lastname: Okhapkina
authors.forename: R O
authors.initials: RO
authors.lastname: Anarbaev
authors.forename: I A
authors.initials: IA
authors.lastname: Lokhova
authors.forename: S K
authors.initials: SK
authors.lastname: Degtyarev
authors.forename: O I
authors.initials: OI
authors.lastname: Lavrik
mesh_terms: D002413:Cations, Divalent; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003854:Deoxyribonucleotides / Q000378:metabolism*; D004795:Enzyme Stability; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D016963:Thermus thermophilus / Q000201:enzymology*; D014961:X-Ray Diffraction
chemical_list: D002413:Cations, Divalent; D003854:Deoxyribonucleotides; D004259:DNA-Directed DNA Polymerase