Shirahata A, Pegg A E
J Biol Chem. 1986 Oct 15;261(29):13833-7.
Total poly(A)-containing mRNA was isolated from rat ventral prostate and translated in a reticulocyte lysate system. The proteins corresponding to S-adenosylmethionine decarboxylase were precipitated with a specific antiserum to this protein. Two proteins were found; one having an Mr of 32,000, which corresponded to the subunit of this enzyme, and a larger protein of Mr 37,000. Immunopurification of polysomes with the antiserum to S-adenosylmethionine decarboxylase followed by mRNA extraction yielded an mRNA preparation which was 10-30% pure mRNA for S-adenosylmethionine decarboxylase. The translation of this mRNA showed clearly that the protein of Mr 37,000 was a precursor of the Mr 32,000 S-adenosylmethionine decarboxylase subunit. Treatment with 2-difluoromethylornithine, which depletes cellular spermidine and is known to increase the content of S-adenosylmethionine decarboxylase protein, led to a 9-fold increase in the content of its mRNA, but treatment with methylglyoxal bis(guanylhydrazone) did not change the mRNA content.
从大鼠腹侧前列腺中分离出总含聚腺苷酸的mRNA,并在网织红细胞裂解物系统中进行翻译。用针对该蛋白的特异性抗血清沉淀与S-腺苷甲硫氨酸脱羧酶相对应的蛋白质。发现了两种蛋白质;一种分子量为32,000,对应于该酶的亚基,另一种分子量较大的蛋白质为37,000。用针对S-腺苷甲硫氨酸脱羧酶的抗血清对多核糖体进行免疫纯化,然后提取mRNA,得到一种mRNA制剂,其纯度为S-腺苷甲硫氨酸脱羧酶的10 - 30%。该mRNA的翻译清楚地表明,分子量为37,000的蛋白质是分子量为32,000的S-腺苷甲硫氨酸脱羧酶亚基的前体。用2-二氟甲基鸟氨酸处理可耗尽细胞内的亚精胺,已知该处理会增加S-腺苷甲硫氨酸脱羧酶蛋白的含量,结果导致其mRNA含量增加9倍,但用甲基乙二醛双(脒腙)处理并未改变mRNA含量。