Rechkunova N I, Okhapkina S S, Anarbaev R O, Lokhova I A, Degtyarev S K, Lavrik O I
Novosibirsk Institute of Bioorganic Chemistry, Siberian Branch of the Russian Academy of Sciences, Novosibirsk.
Biochemistry (Mosc). 2000 May;65(5):609-14.
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.
研究了嗜热栖热菌B35（Tte-pol）的DNA聚合酶在不同二价金属离子存在下与脱氧核苷三磷酸的相互作用。用相应的动力学方案描述了DNA合成以及非互补dNTP对聚合酶反应的竞争性抑制。研究了一些金属（Mg2+、Mn2+、Co2+、Ni2+、Cu2+、Ca2+、Cd2+和Zn2+）的辅因子特性，并确定了它们的激活浓度范围。发现当Mn2+取代Mg2+时，kcat值显著降低，Km值缓慢降低。DNA模板引物的Kd值与Me2+无关，而非互补dNTP的Kd值在Mn2+存在时降低。Tte-pol的持续合成能力而非DNA合成效率与Me2+类型无关。
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