Miyakawa I, Tokitaka M, Shiiba D, Sawada T, Fujikura Y, Fukumoto T, Sando N
Biological Institute, Faculty of Science, Yamaguchi University, Japan.
Plant Cell Physiol. 1993 Jan;34(1):151-6.
Monoclonal antibodies (mAbs) were raised against yeast mitochondrial nucleoids (mt-nucleoids). In an analysis by a combination of immunofluorescence microscopy and staining with 4',6-diamidino-2-phenylindole (DAPI), one of them, designated YMN-1, distinctly stained mt-nucleoids, which were visible as dots, in spheroplasts and in isolated mitochondria. However, staining of isolated mt-nucleoids was rather weak. YMN-1 mAb recognized a 48-kDa protein in immunoblots of both mitochondrial and mt-nucleoid proteins. The 48-kDa protein was a minor component of mt-nucleoid proteins and was separated from extract of both mitochondria and mt-nucleoids by immunoaffinity chromatography. The affinity-purified 48-kDa protein reassociated with mt-nucleoids when mixed with isolated mt-nucleoids, as monitored by immunofluorescence microscopy. The results suggest that a large amount of 48-kDa protein is associated with mt-nucleoids in vivo, and that lysis of mitochondria by the treatment with detergent releases a considerable amount of this protein from mt-nucleoids during the isolation of mt-nucleoids.
制备了针对酵母线粒体类核（mt - 类核）的单克隆抗体（mAb）。在通过免疫荧光显微镜和4',6 - 二脒基 - 2 - 苯基吲哚（DAPI）染色相结合的分析中，其中一种名为YMN - 1的抗体能清晰地对mt - 类核进行染色，在原生质球和分离的线粒体中，mt - 类核呈点状可见。然而，对分离的mt - 类核的染色相当弱。YMN - 1单克隆抗体在线粒体和mt - 类核蛋白的免疫印迹中识别出一种48 kDa的蛋白质。该48 kDa蛋白质是mt - 类核蛋白的次要成分，通过免疫亲和层析从线粒体和mt - 类核提取物中分离出来。通过免疫荧光显微镜监测，当与分离的mt - 类核混合时，亲和纯化的48 kDa蛋白质会与mt - 类核重新结合。结果表明，大量的48 kDa蛋白质在体内与mt - 类核相关联，并且在分离mt - 类核的过程中，用去污剂处理线粒体导致其裂解，从而使大量这种蛋白质从mt - 类核中释放出来。
