Strocchi P, Gilbert J M, Marotta C A
J Neurochem. 1981 Jun;36(6):2044-51. doi: 10.1111/j.1471-4159.1981.tb10832.x.
Studies were undertaken to determine whether there exist variations among the translation products of polysomes from different brains of animals of the same strain. Polysomes were prepared from individual rat cortices and translated in a reticulocyte protein-synthesizing system containing rabbit reticulocyte factors and L-[35S]methionine; he resulting radioactive proteins were analyzed by two-dimensional polyacrylamide gel electrophoresis autoradiography. Comparison of the autoradiographs revealed that two acidic proteins, A and B, of apparent 54,000 M. W. occur as three phenotype: A only, B only, or A plus B. These proteins were not detectable by Coomassie brilliant blue staining of two-dimensional electrophoretograms of brain protein preparations. Messenger RNA was extracted from pooled cortices and translated in a wheat germ extract, and both A and B proteins were detected among the products of translation. Cyclic AMP affinity chromatography of the translation products of cortical polysomes showed that both A and B proteins bind to cyclic AMP. Our data are consistent with the conclusion that there are qualitative differences in the polysome translation products that bind to cyclic AMP among individual cortices of rats of the same strain.
开展了多项研究,以确定同一品系动物不同大脑来源的多核糖体翻译产物之间是否存在差异。从单个大鼠皮质制备多核糖体,并在含有兔网织红细胞因子和L-[35S]甲硫氨酸的网织红细胞蛋白质合成系统中进行翻译;通过二维聚丙烯酰胺凝胶电泳放射自显影分析所得的放射性蛋白质。放射自显影片的比较显示,两种表观分子量为54,000的酸性蛋白质A和B呈现出三种表型:仅A、仅B或A加B。在脑蛋白质制备物的二维电泳图谱中,用考马斯亮蓝染色无法检测到这些蛋白质。从汇集的皮质中提取信使核糖核酸,并在小麦胚芽提取物中进行翻译,在翻译产物中检测到了A和B两种蛋白质。皮质多核糖体翻译产物的环磷酸腺苷亲和层析表明,A和B两种蛋白质都能与环磷酸腺苷结合。我们的数据与以下结论一致,即同一品系大鼠个体皮质中与环磷酸腺苷结合的多核糖体翻译产物存在质的差异。