Affinity labelling of Escherichia coli ribosomes with a benzylidene derivative of AUGU6 within initiation and pretranslocational complexes

FEBS Letters

Karpova, G.G.²  Gren, E.J.¹  Vladimirov, S.N.²  Babkina, G.T.²  Berzin, V.A.¹  Cielens, I.E.²  Yamkovoy, V.I.²  Veniaminova, A.G.² 

[1] Institute of Organic Synthesis, Academy of Sciences of the Latvian SSR, Riga, USSR;Institute of Bioorganic Chemistry, Siberian Division of the USSR Academy of Sciences, Novosibirsk 630090 USSR

关键词: Affinity labeling;    Ribosome;    Ribosomal protein;    mRNA-binding site;    Initiation complex;    Oligonucleotide derivative;    Pretranslocational complex;   

DOI  :  10.1016/0014-5793(86)80714-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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-tRNA_f ^Met and initiation factors and within the pretranslocational complex (complex II) obtained by treatment of complex I with the ternary complex Phe-tRNA^Phe-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-tRNA_f ^Met.

【 授权许可】

Unknown