Lambert P P, Lammens-Verslijpe M, Sennesael J, Herremans N, Delsaux B
Queen Elisabeth Medical Foundation, Brussels, Belgium.
Biol Reprod. 1987 Nov;37(4):887-99. doi: 10.1095/biolreprod37.4.887.
This paper reports the separation of highly cationic proteins (i.e. pI greater than 9.0) of bovine allantoic fluid and their possible pathogenic properties. Experimentally, polycations and cationic proteins of pI greater than 10 induced intravascular coagulation and hemolysis, as well as precipitation of fibrinogen and proteinuria. Bovine allantoic fluid collected at the time of calving contains from 0.6 to 1.3 g of proteins per liter (11 samples). Ion-exchange chromatography, followed by either chromatofocusing or heparin-sepharose-6 beta binding, and, finally, gel filtration separated several fractions and subfractions. These were examined later using polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, and at least ten constituents were visualized. Two components, Mr 34,000, pI 9.7, and Mr 38,000, pI 9.6-9.0, accounted for 33% of the basic proteins present in allantoic fluid and 0.7% of its total protein content. Electrophoretic mobility was unaltered by beta-mercaptoethanol, and periodic acid-Schiff staining was negative. These proteins were not found in ox plasma. The major basic proteins were bound to red cells and platelets. Cell electrophoretic mobility decreased linearly with the logarithm of protein concentration. At concentrations between 10(-6) and 10(-5) M, red cell clumping was rare; hemolysis and platelet agglutination were not observed.
本文报道了牛尿囊液中高阳离子蛋白（即等电点大于9.0）的分离及其可能的致病特性。实验中，等电点大于10的聚阳离子和阳离子蛋白可诱导血管内凝血和溶血，以及纤维蛋白原沉淀和蛋白尿。产犊时收集的牛尿囊液每升含有0.6至1.3克蛋白质（11个样本）。通过离子交换色谱，随后进行色谱聚焦或肝素-琼脂糖-6β结合，最后通过凝胶过滤分离出几个级分和亚级分。随后在十二烷基硫酸钠存在下使用聚丙烯酰胺凝胶电泳对这些进行检查，至少有十种成分可见。两种成分，分子量34,000，等电点9.7，以及分子量38,000，等电点9.6 - 9.0，占尿囊液中碱性蛋白的33%及其总蛋白含量的0.7%。β-巯基乙醇不改变电泳迁移率，过碘酸-希夫染色为阴性。这些蛋白在牛血浆中未发现。主要碱性蛋白与红细胞和血小板结合。细胞电泳迁移率随蛋白质浓度的对数呈线性下降。在10^(-6)至10^(-5) M的浓度之间，红细胞聚集很少见；未观察到溶血和血小板凝集。
