Shibata S, Yokoyama M
Clinical Research Institute, National Medical Center, Tokyo, Japan.
Nephron. 1990;55(2):152-8. doi: 10.1159/000185944.
Normal rat renal cortex contains three renal disease-inducing substances, that is, nephritogenoside, FX1A and immunogenoside (a novel renal glycoprotein that has the biological activity to induce only active Heymann nephritis). When renal cortex homogenate was treated with a detergent, Triton X-100, nephritogenoside was not eluted in the supernatant in which massive doses of FX1A and immunogenoside were detected. Thus, we searched for nephritogenoside in the precipitate. The precipitate was solubilized with trypsin, and nephritogenoside was easily and effectively isolated through columns of DEAE-cellulose followed by Bio-Gel A-1.5 m. The purified sample thus obtained is only composed of nephritogenoside, and contains neither FX1A nor immunogenoside. The yield of purified nephritogenoside was 3.8 mg (starting from 150 rats), which is about 5-6 times the recovery of the previous methodology. This newly developed simple method may be useful for the isolation of purified nephritogenoside. The chemical and immunologic properties of the purified sample prepared by the new method corresponded well with those of the nephritogenoside which was obtained by the previous methodology after exhaustive digestions with three proteolytic enzymes from homologous renal cortex.
正常大鼠肾皮质含有三种可诱发肾病的物质，即肾炎糖苷、FX1A和免疫糖苷（一种新型肾糖蛋白，仅具有诱导活动性海曼肾炎的生物活性）。当用去污剂Triton X-100处理肾皮质匀浆时，肾炎糖苷未在上清液中洗脱出来，而上清液中检测到大量的FX1A和免疫糖苷。因此，我们在沉淀物中寻找肾炎糖苷。沉淀物用胰蛋白酶溶解，然后通过DEAE-纤维素柱，再经Bio-Gel A-1.5m柱，轻松有效地分离出肾炎糖苷。由此获得的纯化样品仅由肾炎糖苷组成，既不含FX1A也不含免疫糖苷。纯化的肾炎糖苷产量为3.8mg（从150只大鼠开始），约为先前方法回收率的5至6倍。这种新开发的简单方法可能有助于分离纯化的肾炎糖苷。用新方法制备的纯化样品的化学和免疫学性质与先前方法从同源肾皮质经三种蛋白水解酶彻底消化后获得的肾炎糖苷的性质非常吻合。
