Necessity of detergent for efficient puromycin-mediated release of nascent peptides from rat liver ribosomes.
Necessity of detergent for efficient puromycin-mediated release of nascent peptides from rat liver ribosomes.
复制标题
需要使用去垢剂来有效地介导嘌呤霉素介导的大鼠肝核糖体中的新生肽的释放。
DOI：
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发表时间：
1981
期刊：
影响因子：
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通讯作者：
T. Omura
中科院分区：
文献类型：
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作者：
T. Oda;N. Nabi;T. Omura
嘌呤霉素介导的新生肽从大鼠肝核糖体的体外释放显着刺激低浓度的洗涤剂的存在下，和刺激是更显着的结合核糖体比游离核糖体。新生肽从核糖体的释放可以分两步进行，首先在不存在去污剂的情况下用嘌呤霉素，然后用去污剂，分别得到两个单独的新生肽级分S1和S2。虽然S1和S2馏分的疏水性和嘌呤霉素结合肽的大小没有显着差异时，通过烷基-琼脂糖柱色谱法和SDS-聚丙烯酰胺凝胶电泳检查，通过免疫沉淀分离的释放肽显示出显着差异的新生肽的两个特定的蛋白质，血清白蛋白和NADPH-细胞色素c还原酶，这两个馏分之间的分布。血清白蛋白的新生肽主要存在于结合核糖体的组分S1中。另一方面，在游离核糖体的组分S2中检测到的NADPH-细胞色素c还原酶的新生肽的比例比其他组分中的大。去污剂的存在下是必不可少的有效的新生肽从核糖体的嘌呤霉素体外释放，这一发现可能是重要的研究在哺乳动物细胞中的特定蛋白质的合成。
Puromycin-mediated in vitro release of nascent peptides from rat liver ribosomes was significantly stimulated by the presence of low concentrations of a detergent, and the stimulation was much more marked with bound ribosomes than with free ribosomes. The release of nascent peptides from ribosomes could be carried out in two steps, first with puromycin in the absence of a detergent and then with a detergent, to give two separate nascent peptide fractions S1 and S2, respectively. Although S1 and S2 fractions were not significantly different in hydrophobicity and in the size of the puromycin-conjugated peptides when examined by alkyl-Sepharose column chromatography and SDS-polyacrylamide gel electrophoresis, the fractionation of the released peptides by immunoprecipitation showed significant difference in the distribution of the nascent peptides of two specific proteins, serum albumin and NADPH-cytochrome c reductase, between these two fractions. The nascent peptides of serum albumin were found mainly in fraction S1 obtained from bound ribosomes. On the other hand, a larger portion of the nascent peptides of NADPH-cytochrome c reductase was detected in fraction S2 from free ribosomes than in other fractions. The presence of a detergent is indispensable for efficient in vitro release of nascent peptides from ribosomes by puromycin and this finding may be important in studying the synthesis of specific proteins in mammalian cells.