蛋白质合成所需的所有因子都保留在与琼脂糖结合的肝素上。
All factors required for protein synthesis are retained on heparin bound to Sepharose.
作者信息
Hradec J, Dusek Z
出版信息
Biochem J. 1978 Apr 15;172(1):1-7. doi: 10.1042/bj1720001.
Abstract
- Postmitochondrial supernatants of rabbit reticulocyte lysates were chromatographed on heparin bound to Sepharose 4B, and the fraction retained on affinity columns was separated by subsequent gel filtration on Sepharose 4B into three fractions, two of them active in protein synthesis. 2. The heavier fraction sedimented at 40S and contained more than 10% RNA. This consisted predominantly of a 12S component, with smaller amounts of the 9S and 4S RNA species. The lighter fraction (18-20S) was composed of proteins with less than 1% RNA. 3. Different enzymic activities were associated with these fractions. 4. In the presence of both fractions, efficient translation took place on combined ribosomal subunits of rat liver with added cofactors. Globin messenger ribonucleoprotein stimulated this translation 5-6-fold. 5. Relatively large complexes of all factors required for protein synthesis are apparently isolated from reticulocytes by affinity chromatography on heparin-Sepharose 4B. Such complexes may occur naturally in the cytoplasm of mammalian cells.
摘要
- 将兔网织红细胞裂解物的线粒体后上清液在结合于琼脂糖4B的肝素上进行层析，然后通过在琼脂糖4B上的后续凝胶过滤，将保留在亲和柱上的组分分离成三个组分，其中两个在蛋白质合成中具有活性。2. 较重的组分在40S处沉降，含有超过10%的RNA。这主要由一个12S组分组成，还有少量的9S和4S RNA种类。较轻的组分（18 - 20S）由RNA含量低于1%的蛋白质组成。3. 不同的酶活性与这些组分相关。4. 在两种组分都存在的情况下，添加辅因子后，大鼠肝脏的核糖体亚基组合上发生了高效翻译。珠蛋白信使核糖核蛋白刺激这种翻译5 - 6倍。5. 蛋白质合成所需的所有因子的相对大的复合物显然通过肝素 - 琼脂糖4B上的亲和层析从网织红细胞中分离出来。这种复合物可能天然存在于哺乳动物细胞的细胞质中。
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