Amara S G, Rosenfeld M G, Birnbaum R S, Roos B A
J Biol Chem. 1980 Apr 10;255(7):2645-8.
Messenger RNA from calcitonin-secreting rat medullary thyroid carcinoma lines directs the synthesis of a putative precursor of calcitonin when translated in wheat embryo lysate. The major translation product directed by mRNA isolated from a rat tumor with a calcitonin content representing greater than 2% of the total protein mass has an apparent molecular weight of 17,500 when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunoprecipitations of the cell-free translation products demonstrate that the 17,500-dalton protein has immunologic determinants which are specifically recognized by antisera to calcitonin. In addition, the relative prominence of this protein is significantly diminished when mRNA from lines with low calcitonin content is used to direct cell-free protein synthesis. This evidence suggests that the initial translation product of calcitonin mRNA is a 17,500-dalton species which must undergo subsequent processing to generate the secreted polypeptide.
来自分泌降钙素的大鼠甲状腺髓样癌细胞系的信使核糖核酸(mRNA),在小麦胚裂解物中进行翻译时,可指导降钙素假定前体的合成。从降钙素含量占总蛋白量2%以上的大鼠肿瘤中分离出的mRNA所指导的主要翻译产物,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析,其表观分子量为17,500。对无细胞翻译产物进行免疫沉淀表明,17,500道尔顿的蛋白质具有免疫决定簇,能被抗降钙素抗血清特异性识别。此外,当使用降钙素含量低的细胞系的mRNA来指导无细胞蛋白质合成时,这种蛋白质的相对丰度会显著降低。这一证据表明,降钙素mRNA的初始翻译产物是一种17,500道尔顿的物质,它必须经过后续加工才能产生分泌型多肽。