Reyes R, Vazquez D, Ballesta J P
Biochim Biophys Acta. 1976 Jul 16;435(4):317-32. doi: 10.1016/0005-2787(76)90198-2.
80-S ribosomes and 60-S subunits from rat liver were treated at increasing KC1 concentrations giving protein-deficient ribosomal particles whose components were analyzed and their activity tested. Most of the activities assayed stand treatment up to KC1 concentrations of around 0.6 M; peptidyl transferase, measured by the fragment reaction, however was 50% inhibited by 0.5 M KC1 in 60-S subunits but not in 80-S ribosomes. Three proteins, L21, L26 and L31, might be implicated in this loss of activity. 60-S subunits forming part of the 80 S ribosome are more resistant to the salt treatment and the pattern of proteins released by the treatment differs from the one obtained from free 60-S subunits, implying perhaps a change of conformation of this subunit upon association to form 80-S couples. According to their resistance to release by KC1 the proteins of the large sub-unit can be divided into three groups: (1) easily removed, including proteins: L1, L11, L17 and L25 in 80-s subunits and in addition, L5, L8, L9, L13, L20, L22, L26, L29, L31 and L32/33 in 60-S subunits; (2) proteins resistant to release by high salt concentrations in 80-S ribosomes as well as in 60-S subunits, namely proteins L3, L14, L27, L36, L40, L41, X1 and X2; (3) the rest of the proteins which are released in a more or less continuous way throughout the treatment. 5 S RNA is not released by KC1 treatment at the concentrations used. The binding sites for the antibiotics trichodermin and anisomycin are affected in a different way by the salt treatment, indicating that they are structurally different.
用递增的KCl浓度处理来自大鼠肝脏的80-S核糖体和60-S亚基,得到蛋白质缺乏的核糖体颗粒,对其组分进行分析并测试其活性。所检测的大多数活性在KCl浓度达到约0.6M时仍能耐受处理;然而,通过片段反应测定的肽基转移酶在60-S亚基中被0.5M KCl抑制了50%,但在80-S核糖体中未被抑制。三种蛋白质L21、L26和L31可能与这种活性丧失有关。构成80S核糖体一部分的60-S亚基对盐处理更具抗性,处理释放的蛋白质模式与从游离60-S亚基获得的不同,这可能意味着该亚基在缔合形成80-S对时构象发生了变化。根据它们对KCl释放的抗性,大亚基的蛋白质可分为三组:(1) 容易去除的,包括80-S亚基中的蛋白质L1、L11、L17和L25,此外还有60-S亚基中的L5、L8、L9、L13、L20、L22、L26、L29、L31和L32/33;(2) 在80-S核糖体以及60-S亚基中对高盐浓度释放具有抗性的蛋白质,即蛋白质L3、L14、L27、L36、L40、L41、X1和X2;(3) 在整个处理过程中以或多或少连续的方式释放的其余蛋白质。在所使用的浓度下,5S RNA不会因KCl处理而释放。抗生素曲古抑菌素和茴香霉素的结合位点受盐处理的影响方式不同,表明它们在结构上不同。