Krasnorutskii Michael A, Buneva Valentina N, Nevinsky Georgy A
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, pr. Lavrent'eva 8, Novosibirsk 630090, Russia.
J Mol Recognit. 2008 Jul-Aug;21(4):233-42. doi: 10.1002/jmr.890.
Immunization of rabbits with DNase I leads to the production of antiidiotypic Abs with DNase activity. It is not known at present whether antiidiotypic Abs against DNA-hydrolyzing enzymes can possess RNase activity. Here we show that immunization of healthy rabbits with bovine DNase I produces IgGs with intrinsic DNase and RNase activities. Electrophoretically and immunologically homogeneous polyclonal IgGs were obtained by sequential chromatography of the immune sera on Protein A-Sepharose and gel filtration. Affinity chromatography on DNA cellulose using elution of Abs with different concentrations of NaCl and an acidic buffer separated catalytic IgGs into four Ab subfractions, three of which demonstrated only DNase activity while one subfraction hydrolyzed RNA faster than DNA. The serum of patients with many different autoimmune (AI) diseases contains small fractions of antibodies (Abs) interacting with immobilized DNA, which possess both DNase and RNase activities. Our data suggest that a fraction of abzymes from AI patients hydrolyzing both DNA and RNA can contain a subfraction of Abs against DNase I.
用脱氧核糖核酸酶I免疫兔子会导致产生具有脱氧核糖核酸酶活性的抗独特型抗体。目前尚不清楚针对DNA水解酶的抗独特型抗体是否具有核糖核酸酶活性。在此我们表明,用牛脱氧核糖核酸酶I免疫健康兔子会产生具有内在脱氧核糖核酸酶和核糖核酸酶活性的免疫球蛋白G。通过将免疫血清依次在蛋白A-琼脂糖凝胶上进行层析和凝胶过滤,获得了电泳和免疫均一的多克隆免疫球蛋白G。使用不同浓度的氯化钠和酸性缓冲液洗脱抗体,在DNA纤维素上进行亲和层析,将具有催化活性的免疫球蛋白G分离为四个抗体亚组分,其中三个仅表现出脱氧核糖核酸酶活性,而一个亚组分水解RNA的速度比DNA快。患有多种不同自身免疫性(AI)疾病的患者血清中含有一小部分与固定化DNA相互作用的抗体,这些抗体同时具有脱氧核糖核酸酶和核糖核酸酶活性。我们的数据表明,来自AI患者的一部分同时水解DNA和RNA的抗体酶可能包含针对脱氧核糖核酸酶I的抗体亚组分。