Stafeeva O A, Khaĭlova L S, Katrukha A G, Bulargina T V
Biokhimiia. 1995 Jan;60(1):124-30.
A monoclonal antibody against the E1-component of pigeon breast muscle has been prepared. The dissociation constant of the E1-mAb F7F10 complex was determined to be equal to 5.93.10(-8) M. The cross-reaction between mAb F7F10 and the E1-component of the pyruvate dehydrogenase complex from various species (including human beings) was established. The F7F10 antibody was shown to interact with both alpha- and beta-subunits of E1, which suggests that the amino acid residues in the both subunits are constituents of the antigenic determinant. Binding of the F7F10 antibody to the antigen had no effect on the enzymatic activity of E1 but induced rapid inactivation of the pyruvate dehydrogenase complex in the pyruvate:NAD oxidoreductase reaction. The competition between the F7F10 antibody and the E2-component of the pyruvate dehydrogenase complex for the binding to E1 was revealed by immunoenzymatic analysis. It was concluded that the antigenic determinant and the E1 site responsible for the E1-E2 interaction within the pyruvate dehydrogenase complex may overlap.
已制备出一种针对鸽胸肌E1成分的单克隆抗体。E1-单克隆抗体F7F10复合物的解离常数经测定等于5.93×10⁻⁸ M。确定了单克隆抗体F7F10与来自各种物种（包括人类）的丙酮酸脱氢酶复合物的E1成分之间存在交叉反应。F7F10抗体显示能与E1的α亚基和β亚基相互作用，这表明两个亚基中的氨基酸残基都是抗原决定簇的组成部分。F7F10抗体与抗原的结合对E1的酶活性没有影响，但在丙酮酸：NAD氧化还原酶反应中可诱导丙酮酸脱氢酶复合物快速失活。免疫酶分析揭示了F7F10抗体与丙酮酸脱氢酶复合物的E2成分在与E1结合上存在竞争。得出的结论是，丙酮酸脱氢酶复合物内的抗原决定簇与负责E1-E2相互作用的E1位点可能重叠。
