Zeichner-David M, Weliky B G, Slavkin H C
Biochem J. 1980 Feb 1;185(2):489-96. doi: 10.1042/bj1850489.
Experiments were designed to identify and characterize tissue-specific proteins involved in the process of tooth organogenesis. Epithelial and mesenchymal proteins were extracted from intact molar organs or mechanically separated tissues obtained from 25-day New Zealand White rabbit embryos. Labelling experiments with [35S]methionine followed by radioautography or gel electrophoresis and fluorography showed the presence of label only in epithelial proteins. Most of these proteins range from 43 000 mol.wt. and higher, except for one band of approx. 16 000 mol.wt. A mRNA fraction of 16--26S was isolated by ultracentrifugation on sucrose gradients. When translated in a reticulocyte-lysate cell-free system, the mRNA obtained from intact molar organs resulted in the synthesis of three proteins, of mol.wts. 65 000, 58 000 and 43 000. A similar mRNA fraction obtained from dental-pulp mesenchyme gave only the 43 000-mol.wt. protein, indicating that the 65 000- and 58 000-mol.wt. proteins are derived from epithelial cells.
实验旨在鉴定和表征参与牙齿器官形成过程的组织特异性蛋白质。从25日龄新西兰白兔胚胎的完整磨牙器官或机械分离的组织中提取上皮和间充质蛋白质。用[35S]甲硫氨酸进行标记实验,然后进行放射自显影或凝胶电泳和荧光自显影,结果表明标记仅存在于上皮蛋白质中。这些蛋白质大多数分子量在43000及以上,除了一条约16000分子量的条带。通过在蔗糖梯度上超速离心分离出16 - 26S的mRNA组分。当在网织红细胞裂解物无细胞系统中进行翻译时,从完整磨牙器官获得的mRNA导致合成了三种蛋白质,分子量分别为65000、58000和43000。从牙髓间充质获得的类似mRNA组分只产生了分子量为43000的蛋白质,这表明分子量为65000和58000的蛋白质来自上皮细胞。