在起始复合物和转位前复合物中用AUGU6的亚苄基衍生物对大肠杆菌核糖体进行亲和标记。
Affinity labelling of Escherichia coli ribosomes with a benzylidene derivative of AUGU6 within initiation and pretranslocational complexes.
作者信息
Babkina G T, Veniaminova A G, Vladimirov S N, Karpova G G, Yamkovoy V I, Berzin V A, Gren E J, Cielens I E
出版信息
FEBS Lett. 1986 Jul 7;202(2):340-4. doi: 10.1016/0014-5793(86)80714-1.
Affinity labelling of E. coli ribosomes with the 2',3'-O-[4-(N-2-chloroethyl)-N-methylamino]benzylidene derivative of AUGU6 was studied within the initiation complex (complex I) obtained by using fMet-tRNAMetf and initiation factors and within the pretranslocational complex (complex II) obtained by treatment of complex I with the ternary complex Phe-tRNAPhe.GTP.EF-Tu. Both proteins and rRNA of 30 S as well as 50 S subunits were found to be labelled. Sets of proteins labelled within complexes I and II differ considerably. Within complex II, proteins S13 and L10 were labelled preferentially. On the other hand, within complex I, multiple modification is observed (proteins S4, S12, S13, S14, S15, S18, S19, S20/L26 were found to be alkylated) despite the single fixation of a template in the ribosome by interaction of the AUG codon with fMet-tRNAMetf.
利用甲酰甲硫氨酰 - tRNAMetf和起始因子获得起始复合物（复合物I），并用三元复合物苯丙氨酰 - tRNAPhe.GTP.EF - Tu处理复合物I获得转位前复合物（复合物II），研究了用AUGU6的2',3'-O-[4-(N - 2 - 氯乙基)-N - 甲基氨基]亚苄基衍生物对大肠杆菌核糖体进行亲和标记的情况。结果发现30 S和50 S亚基的蛋白质和rRNA均被标记。在复合物I和复合物II中被标记的蛋白质组有很大差异。在复合物II中，蛋白质S13和L10被优先标记。另一方面，在复合物I中，尽管AUG密码子与甲酰甲硫氨酰 - tRNAMetf相互作用使模板在核糖体中单一固定，但仍观察到多个修饰（发现蛋白质S4、S12、S13、S14、S15、S18、S19、S20/L26被烷基化）。
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