Muno D, Kominami E, Ishii H, Usui K, Saifuku K, Sakakibara Y, Namihisa T
Department of Biochemistry, Juntendo University, School of Medicine, Tokyo, Japan.
Hepatology. 1990 Jan;11(1):16-23. doi: 10.1002/hep.1840110105.
Most of the sera from patients with primary biliary cirrhosis contains antimitochondrial antibodies, which react with four proteins of the mitochondrial inner membrane. We reported in a previous paper that when beef heart mitochondrial inner membrane proteins were digested by trypsin, a new reactive 36 kDa fragment with antimitochondrial antibody was obtained. This 36 kDa fragment derives from original 70 kDa protein because the monoclonal antibody specific to 70 kDa protein reacts with the 36 kDa band equivalent to 70 kDa band. The 36 kDa fragment was purified using an affinity column conjugated with an immunoglobulin-rich fraction of primary biliary cirrhosis serum containing antimitochondrial antibody, preparative electrophoresis and high-performance liquid chromatography using a reverse phase column. The final preparation showed a single band in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Its amino acid composition is in good agreement with that of the subunit binding domain of the pyruvate dehydrogenase complex E2 from bovine heart.
大多数原发性胆汁性肝硬化患者的血清中含有抗线粒体抗体，这些抗体可与线粒体内膜的四种蛋白质发生反应。我们在之前的一篇论文中报道，当牛心线粒体内膜蛋白用胰蛋白酶消化时，可获得一种与抗线粒体抗体反应的新的36 kDa活性片段。这个36 kDa片段源自原来的70 kDa蛋白，因为特异性针对70 kDa蛋白的单克隆抗体可与相当于70 kDa条带的36 kDa条带发生反应。使用与含有抗线粒体抗体的原发性胆汁性肝硬化血清的富含免疫球蛋白部分偶联的亲和柱、制备电泳以及使用反相柱的高效液相色谱法对36 kDa片段进行纯化。最终制剂在十二烷基硫酸钠聚丙烯酰胺凝胶电泳中显示为单一一条带。其氨基酸组成与牛心丙酮酸脱氢酶复合体E2的亚基结合结构域的氨基酸组成高度一致。
