Tejler L, Grubb A O
Biochim Biophys Acta. 1976 Jul 19;439(1):82-94. doi: 10.1016/0005-2795(76)90164-1.
A glycoprotein from the urine of one healthy individual was purified by ultrafiltration, ion-exchange chromatography, gel chromatography and immunosorption. The protein contained only one polypeptide chain with an approximate molecular weight of 31 000 and was associated with a brown colour which did not disappear even after total reduction and alkylation of the protein followed by dialysis in 6 M guanidine hydrochloride. The protein appeared homogeneous on sodium dodecyl sulphate-polyacrylamide electrophoresis and gel chromatography. It had only one N-terminal amino acid sequence, Gly-Pro, and gave only one precipitate with a polyvalent antiserum but was found to be very heterorgeneous on agarose gel electrophoresis and on isoelectric focusing. Desialylation of the protein failed to alter this heterogeneity. An electroimmunoassay system was designed to measure the amount of the protein in normal human plasma, urine, and cerebrospinal fluid where the mean concentrations were found to be about 100, 10 and 0.3 mg/l, respectively. The protein was found to occur in normal plasma and urine as free monomers and dimers and as complexes with IgA and albumin.
从一名健康个体的尿液中提取的一种糖蛋白,通过超滤、离子交换色谱、凝胶色谱和免疫吸附等方法进行了纯化。该蛋白质仅含有一条多肽链,分子量约为31000,且呈现出棕色,即使在蛋白质完全还原和烷基化后,于6M盐酸胍中透析,这种棕色也不会消失。该蛋白质在十二烷基硫酸钠-聚丙烯酰胺电泳和凝胶色谱上显示为均一状态。它只有一个N端氨基酸序列,即甘氨酸-脯氨酸,与多价抗血清反应时只产生一种沉淀,但在琼脂糖凝胶电泳和等电聚焦中发现其非常不均一。对该蛋白质进行脱唾液酸处理未能改变这种不均一性。设计了一种电免疫分析系统来测定正常人血浆、尿液和脑脊液中该蛋白质的含量,发现其平均浓度分别约为100mg/L、10mg/L和0.3mg/L。发现该蛋白质在正常血浆和尿液中以游离单体和二聚体形式存在,也以与IgA和白蛋白的复合物形式存在。