Structural and Functional Properties of Ribosomes Crosslinked with Dimethylsuberimidate

AUTOR(ES)

Slobin, Lawrence I.

RESUMO

To test whether a 30S ribosomal subunitformylmethionyl-tRNA-mRNA complex is an obligatory intermediate in protein synthesis, 70S ribosomes from Escherichia coli were crosslinked with the bifunctional imidoester, dimethylsuberimidate. Crosslinked ribosomes contained covelently joined 30S and 50S subunits, as judged by their inability to dissociate at low Mg2+ concentrations. Treatment of 70S ribosomes with high salt (1 M NH4Cl), either before or after reaction with the crosslinking reagent, produced two different crosslinked ribosomal particles, one of “60 S” and the other “70 S.” Preliminary evidence indicates that both particles can bind N-acetylphenylalanyl-tRNA at low Mg2+ concentrations and are active for polyphenylalanine syntheses.

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