Dzau V J, Brenner A, Wolfsohn S, Haber E
Hypertension. 1982 May-Jun;4(3):341-7. doi: 10.1161/01.hyp.4.3.341.
Antibodies raised to pure canine renal renin were used to probe homology of renin from other species. Goat, rabbit, and mouse antibodies exhibited similar properties and were specific for renin as confirmed by immunodiffusion, immunoelectrophoresis, and selective inhibition of renin enzymatic activity. Goat antibody also inhibited the enzymatic activities of rat, hog, and bovine renin. Immunologic cross reactivity was further confirmed by direct binding and competition assays. Goat anti-(canine renin) antibody did not inhibit human or mouse renin activity, but readily bound to these enzymes, suggesting the presence of epitopes distant from the enzymatic site. Purified goat Fab retained only 10% to 20% of the anticatalytic activity of antibody, but this activity was largely recovered when donkey anti-goat antibody was added to the Fab-renin mixture. Thus, the major fraction (80% to 90%) of goat antibody exerted its anticatalytic activity by immune complex formation of steric hindrance and is not catalytic-site directed, whereas 10% to 20% binds an epitope at or near the catalytic site.
用针对纯犬肾素制备的抗体来探究其他物种肾素的同源性。山羊、兔子和小鼠的抗体表现出相似的特性,且通过免疫扩散、免疫电泳以及肾素酶活性的选择性抑制证实对肾素具有特异性。山羊抗体还抑制大鼠、猪和牛肾素的酶活性。免疫交叉反应通过直接结合和竞争试验进一步得到证实。山羊抗(犬肾素)抗体不抑制人或小鼠肾素活性,但能与这些酶轻松结合,这表明存在远离酶活性位点的表位。纯化的山羊Fab片段仅保留了抗体10%至20%的抗催化活性,但当将驴抗山羊抗体添加到Fab-肾素混合物中时,该活性大部分得以恢复。因此,山羊抗体的主要部分(80%至90%)通过空间位阻的免疫复合物形成发挥其抗催化活性,并非针对催化位点,而10%至20%则结合催化位点或其附近的一个表位。