Tsukada H, Mochizuki Y, Fujiwara S
J Cell Biol. 1966 Mar;28(3):449-60. doi: 10.1083/jcb.28.3.449.
The nucleoids of microbodies of rat liver cells were isolated in a highly homogeneous and pure state, by treating the microbody-rich fraction, prepared from 10% polyvinylpyrrolidone-0.25 M sucrose homogenate, with Triton X-100. Three treatments with 0.1% detergent were enough to render the nucleoids free from contamination with mitochondria, microsomes, lysosomes, and intact microbodies. Electron microscopically, the nucleoids were found to consist of parallel bundles of highly dense hollow tubules, the outer and inner diameters of which are approximately 150 and 50 A, respectively. Ten tubules are arranged around a longitudinal space 190 x 200 A in width. The nucleoids thus show a honeycomb appearance in the cross-plane and a parallel-packed structure in the longitudinal plane. Biochemically, the nucleoids were found to bear only urate oxidase among probably microbody-enzymes, and they might be the only cytoplasmic particles of rat liver cells in which the enzyme locates. Urate oxidase activity, on a unit protein basis, of the nucleoid preparation is approximately 380 times as high as that of the whole homogenate, and is almost comparable with that of a commercial type I enzyme preparation. No enzymes of mitochondrial, microsomal, and lysosomal origins were detected in the nucleoids. The fine structure of the nucleoids is described in detail, and a probable schematic diagram is presented.
通过用Triton X-100处理由10%聚乙烯吡咯烷酮-0.25M蔗糖匀浆制备的富含微体的部分,以高度均一和纯净的状态分离大鼠肝细胞微体的拟核。用0.1%去污剂进行三次处理足以使拟核免受线粒体、微粒体、溶酶体和完整微体的污染。电子显微镜观察发现,拟核由高度密集的中空小管平行束组成,其外径和内径分别约为150和50埃。十根小管围绕着一个宽190×200埃的纵向空间排列。因此,拟核在横切面上呈现蜂窝状外观,在纵切面上呈现平行堆积结构。生化分析发现,在可能的微体酶中,拟核仅含有尿酸氧化酶,并且它们可能是大鼠肝细胞中该酶所在的唯一细胞质颗粒。以单位蛋白计算,拟核制剂的尿酸氧化酶活性约为全匀浆的380倍,几乎与市售I型酶制剂相当。在拟核中未检测到线粒体、微粒体和溶酶体来源的酶。详细描述了拟核的精细结构,并给出了可能的示意图。