Bull P, MacDonald H, Valenzuela P
Biochim Biophys Acta. 1981 May 29;653(3):368-77. doi: 10.1016/0005-2787(81)90193-3.
The interaction between yeast RNA polymerase I and Cibacron blue F3GA has been studied by difference spectrophotometry and column chromatography. The enzyme is reversibly inhibited by the dye. 50% inhibition is obtained with 7.5 x 10(-6) M Cibacron blue. 1 mol Cibacron blue binds per mol enzyme. This interaction, which is inhibited by salt, occurs at a site different from the active site. When RNA polymerase I is chromatographed in Blue dextran-Sepharose columns, two polypeptides, of 48 000 and 36 000 daltons, are dissociated from the enzyme. The resulting enzyme is completely inactive, ATP (5 mM) present in the elution buffer prevents both the dissociation of the polypeptides and the inactivation of the enzyme.
通过差示分光光度法和柱色谱法研究了酵母RNA聚合酶I与汽巴蓝F3GA之间的相互作用。该染料可使酶发生可逆抑制。7.5×10⁻⁶ M汽巴蓝可导致50%的抑制率。每摩尔酶结合1摩尔汽巴蓝。这种相互作用受盐抑制,发生在与活性位点不同的位点。当RNA聚合酶I在蓝色葡聚糖-琼脂糖柱上进行色谱分析时,两种分子量分别为48000和36000道尔顿的多肽从酶上解离下来。所得的酶完全无活性,洗脱缓冲液中存在的5 mM ATP可防止多肽解离和酶失活。