Baronaĭte Z A, Martsishauskas R P, Pesliakas I G
Biokhimiia. 1984 Jan;49(1):142-8.
Using kinetic methods and differential spectrophotometry, the interaction between DNA and RNA ligases T4 and cibacron blue F3GA was studied. It was shown that the dye inhibits the first step of the enzymatic reaction, i. e. the formation of the AMP ligase complex. A 50% inhibition of the AMP-ligase complex by DNA ligase occurs at the dye concentration of 1 X 10(-5) M, that by RNA ligase--at 1 X 10(-4) M. Cibacron blue F3GA also inhibits the formation of end products of the reaction catalyzed by these enzymes. The dye is a noncompetitive inhibitor of RNA ligase with respect to [32P]oligoA20 and a competitive one with respect to ATP. Using differential spectrophotometry, it was found that the interaction occurs predominantly via electrostatic bonds between the SH-groups of the dye and the amino groups of lysine residues. DNA ligase possesses a higher affinity for the dye than RNA ligase.
采用动力学方法和差示分光光度法，研究了DNA与RNA连接酶T4和汽巴克隆蓝F3GA之间的相互作用。结果表明，该染料抑制酶促反应的第一步，即AMP连接酶复合物的形成。DNA连接酶对AMP连接酶复合物产生50%抑制时，染料浓度为1×10⁻⁵ M；RNA连接酶产生50%抑制时，染料浓度为1×10⁻⁴ M。汽巴克隆蓝F3GA还抑制这些酶催化的反应终产物的形成。该染料对[³²P]oligoA₂₀而言是RNA连接酶的非竞争性抑制剂，对ATP而言是竞争性抑制剂。利用差示分光光度法发现，相互作用主要通过染料的SH基团与赖氨酸残基的氨基之间的静电键发生。DNA连接酶对该染料的亲和力高于RNA连接酶。
