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从人胎盘中分离出的致肾炎糖蛋白(从胎盘中分离的方法)

[Nephritogenic glycoprotein isolated from human placenta (methods of isolation from placenta].

作者信息

Iioka H, Moriyama I, Ichijo M, Shibata S

出版信息

Nihon Sanka Fujinka Gakkai Zasshi. 1983 Jun;35(6):827-33.

PMID:6875336
Abstract

In our experiments, we succeeded in obtaining from human placenta a substance comparable to human renal nephritogenic glycoprotein prepared from GBM (glomerular basement membrane). The procedure was quite similar to that for preparation from the kidney. At first, placental terminal villi were isolated by the successive use of three metal sieves. Then trophoblast basement membranes (TrBM) were isolated from placental terminal villi by ultrasonic disruption. Finally TrBM were made soluble by trypsin digestion, after which they were further digested by pronase. Ouchterlony gel diffusion demonstrated the existence of a common precipitin line between antiserum to human renal nephritogenic glycoprotein prepared from GBM and antiserum to the sample prepared from human placental TrBM, and human renal nephritogenic glycoprotein prepared from GBM and the sample prepared from human placental TrBM. The monosaccharide composition of the sample prepared from human placental TrBM was rich in glucose (glucose, galactose and mannoside in the ratio of 1.00 : 0.74 : 1.00), and the amino acid composition of the sample contained no collagenous components. As a result of fractionation of the sample prepared from human placental TrBM by Bio Gel P200 column chromatography, the activity of renal nephritogenic glycoprotein was found within the void fraction.

摘要

在我们的实验中,我们成功地从人胎盘中获得了一种物质,该物质与从肾小球基底膜(GBM)制备的人肾致肾炎糖蛋白相当。该制备过程与从肾脏制备的过程非常相似。首先,通过连续使用三个金属筛分离胎盘终末绒毛。然后通过超声破碎从胎盘终末绒毛中分离滋养层基底膜(TrBM)。最后,通过胰蛋白酶消化使TrBM溶解,之后再用链霉蛋白酶进一步消化。双向免疫扩散试验表明,由GBM制备的人肾致肾炎糖蛋白抗血清与由人胎盘TrBM制备的样品抗血清之间,以及由GBM制备的人肾致肾炎糖蛋白与由人胎盘TrBM制备的样品之间存在共同沉淀线。由人胎盘TrBM制备的样品的单糖组成富含葡萄糖(葡萄糖、半乳糖和甘露糖苷的比例为1.00 : 0.74 : 1.00),且该样品的氨基酸组成不含胶原成分。通过Bio Gel P200柱色谱对由人胎盘TrBM制备的样品进行分级分离后,在空体积部分发现了肾致肾炎糖蛋白的活性。

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