Imada M, Kao F T, Law M L, Jones C
Somat Cell Mol Genet. 1986 Mar;12(2):197-201. doi: 10.1007/BF01560666.
Cell surface proteins and glycoproteins of human and Chinese hamster cells and their hybrid cell clones were analyzed by two-dimensional gel electrophoresis. The J1 clone of human-Chinese hamster hybrid cells contained chromosome 11 as its only human chromosome. The J1 cells expressed a glycoprotein of 200,000 daltons which was shared by human fibroblasts but not by the parental Chinese hamster ovary cells. The 200,000-dalton protein was identified as a cell surface protein by the method of lactoperoxidase-catalyzed iodination. The protein was electrophoretically purified from radioiodinated cultures of human fibroblasts and J1 cells and subjected to the analysis of tryptic peptides by thin-layer electrophoresis followed by chromatography. The protein from both sources gave rise to fingerprints which closely resembled to each other. The results are consistent with a hypothesis that the 200,000-dalton protein of the J1 clone is of human origin. Analysis of segregant clones of J1 cells, which have deletions on human chromosome 11, has further suggested that the gene for this glycoprotein maps to the long arm of chromosome 11. A gene coding for the 200,000-dalton protein has not been previously mapped to this chromosome.
通过二维凝胶电泳分析了人和中国仓鼠细胞及其杂交细胞克隆的细胞表面蛋白和糖蛋白。人-中国仓鼠杂交细胞的J1克隆含有11号染色体,这是其唯一的人类染色体。J1细胞表达一种200,000道尔顿的糖蛋白,人成纤维细胞也表达这种糖蛋白,但其亲本中国仓鼠卵巢细胞不表达。通过乳过氧化物酶催化碘化法将这种200,000道尔顿的蛋白鉴定为细胞表面蛋白。该蛋白从人成纤维细胞和J1细胞的放射性碘化培养物中通过电泳纯化,然后通过薄层电泳继以色谱法对胰蛋白酶肽进行分析。来自两种来源的蛋白产生的指纹图谱彼此非常相似。结果与这样一种假设一致,即J1克隆的200,000道尔顿蛋白是人类起源的。对J1细胞分离克隆的分析进一步表明,这种糖蛋白的基因定位于11号染色体的长臂上,11号染色体长臂上此前尚未定位编码200,000道尔顿蛋白的基因。