Hadding U, Müller-Eberhard H J
Immunology. 1969 Jun;16(6):719-35.
A method is described which permits isolation of C′9 from human serum. The final product was homogeneous by polyacrylamide gel electrophoresis and the distribution of C′9 haemolytic activity corresponded to the position of the protein disc. C′9 is an α-globulin which has a sedimentation coefficient of 4.5S, a diffusion coefficient of 5.1 × 10 cm/sec and a calculated molecular weight of 79,000. The reaction of C′9 with EAC′1∼8 was found to be temperature dependent and to proceed optimally at pH 7. In the course of this reaction C′9 activity declined in the fluid phase and C′9 protein became specifically attached to the cells, as revealed by the use of radioactively labelled C′9. The haemolytic activity of C′9 was mimicked by bipyridine which exhibited the previously reported phenanthroline effect. Kinetic experiments showed that 0.09 M EDTA, which inhibits haemolysis, acts subsequent to the C′9 reaction step. Possible modes of action of C′9 are discussed.
本文描述了一种从人血清中分离C′9的方法。通过聚丙烯酰胺凝胶电泳，最终产物呈均一性，C′9溶血活性的分布与蛋白质条带的位置相对应。C′9是一种α球蛋白，沉降系数为4.5S，扩散系数为5.1×10 cm/秒，计算分子量为79,000。发现C′9与EAC′1∼8的反应依赖于温度，在pH 7时反应最佳。在该反应过程中，C′9活性在液相中下降，并且如使用放射性标记的C′9所显示的，C′9蛋白特异性地附着于细胞。联吡啶模拟了C′9的溶血活性，表现出先前报道的菲咯啉效应。动力学实验表明，抑制溶血的0.09 M EDTA在C′9反应步骤之后起作用。文中讨论了C′9可能的作用方式。
