Identification of the tRNA-binding sites on rat liver ribosomes by affinity labeling

  title={Identification of the tRNA-binding sites on rat liver ribosomes by affinity labeling},
  author={A. P. Czernilofsky and Ekkehard Collatz and Axel M. Gressner and Ira G. Wool and Ernst K{\"u}chler},
  journal={Molecular and General Genetics MGG},
Summaryp-Nitrophenoxycarbonyl-3H-phenylalanyl-tRNAyeastPhebinds to 80S rat liver ribosomes and forms a covalent bond with ribosomal proteins. The affinity labeled proteins were identified by a kind of “three-dimensional” electrophoresis: groups of 80S proteins were cut out of two-dimensional gel slabs, eluted, and separated by electrophoresis in polyacrylamide gels containing sodium dodecyl sulfate; finally, the radioactivity in the protein bands was determined. The proteins L32/L33, L36, L21… 

Identification of proteins at the peptidyl-tRNA binding site of rat liver ribosomes

A model for the arrangement of proteins within this active site of rat liver peptidyl-tRNA is constructed, using a 3′-terminal pentanucleotide fragment of N-acetylleucyl-t RNA in which mercury atoms have been added at the C-5 position of the three cytosine residues.

Affinity labelling of yeast ribosomal peptidyl transferase

When only the components of the ribosomal P-site were analysed by reacting the treated particles with puromycin fewer spots were labelled, corresponding to proteins L36 and L19/20 using PNPC-Phe-tRNA and proteins L4/6, L36, and L43 using N-Iodoacetyl-P he-t RNA.



Proteins at the tRNA binding sites of Escherichia coli ribosomes.

The affinity label is shown to react with the 50S proteins L27, L15, L2, L16, and L14, and it is concluded that these particular proteins are located at or near tRNA-binding sites within the50S ribosomal subunit.

Peptidyl transferase center of rat-liver ribosome cores.

A comparative study of the proteins released by treatment in the two types of particles suggests the involvement in the peptidyl transferase center of the ribosome of one or more of the floolwing proteins: L21, L24, L27, L28 and L36.

Studies on proteins of animal ribosomes. XIII. enumeration of ribosomal proteins of rat liver

Affinity label for the tRNA binding site on the Escherichia coli ribosome.

Protein synthesis initiation in eukaryotes. Characterization of ribosomal factors from mouse fibroblasts.

The initiation complex formed in the sequential system is functional, and under appropriate conditions transfers its methionine to methionyl-puromycin or polypeptide as well as other steps in the sequence are unaffected.

Chemical studies on methionyl-tRNA synthetase from Escherichia coli.