Mitsui K, Nakagawa T, Tsurugi K
Department of Biochemistry, Yamanashi Medical College.
J Biochem. 1988 Dec;104(6):908-11. doi: 10.1093/oxfordjournals.jbchem.a122581.
A small but distinct amount of yeast acidic ribosomal proteins A1/A2 was detected in cytosol by immunoblotting on a two-dimensional gel electrophoretogram, while 38 kDa acidic protein A0 was not detected. The free forms of A1/A2 in the cytosol were eluted in gel filtration at the molecular mass of about 30 kDa under non-denaturation conditions, suggesting that they exist as a dimer or timer without association with A0. The amount of free A1/A2 was determined by immunoblotting to be 0.3% of the ribosome-bound A1/A2 in yeast. The time course of incorporation of radioactive amino acid showed that the cytosolic free A1/A2 are labeled more rapidly with high specific radioactivity than the ribosome-bound A1/A2. This result suggested that some of the cytosolic A1/A2, if not all, are newly-synthesized proteins which are ready for incorporation into cytoplasmic ribosomes.
通过二维凝胶电泳图上的免疫印迹法在细胞溶胶中检测到少量但明显的酵母酸性核糖体蛋白A1/A2,而未检测到38 kDa的酸性蛋白A0。在非变性条件下,细胞溶胶中A1/A2的游离形式在凝胶过滤中以约30 kDa的分子量被洗脱,这表明它们以二聚体或四聚体形式存在,且不与A0结合。通过免疫印迹法确定游离A1/A2的量为酵母中核糖体结合型A1/A2的0.3%。放射性氨基酸掺入的时间进程表明,细胞溶胶中的游离A1/A2比核糖体结合型A1/A2更快地被高比放射性标记。这一结果表明,部分(如果不是全部)细胞溶胶中的A1/A2是新合成的蛋白质,随时准备掺入细胞质核糖体。