Molecular and General Genetics MGG

, Volume 153, Issue 3, pp 231–235 | Cite as

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

  • A. Peter Czernilofsky
  • Ekkehard Collatz
  • Axel M. Gressner
  • Ira G. Wool
  • Ernst Küchler
Article

Summary

p-Nitrophenoxycarbonyl-3H-phenylalanyl-tRNA yeast Phe binds 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, L23, L28/L29 and L13 were labeled when the reaction was with poly(U) present. We conclude they are located at or near the peptidyl transferase center. Proteins S7 and/or L9 and a protein in the vicinity of L36 which could not be identified were more radioactive when the affinity reaction was in the absence of poly(U).

Abbreviations

PNPC-Phe-tRNA

p-nitrophenoxycarbonyl-3H-phenylalanyl-tRNA yeast Phe (phenylalanine specific, isolated from yeast)

HEPES

N-2-hydroxyethylpiperazine-N′-2-ethane sulfonic acid

PPO

2,5-diphenyloxazol

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Copyright information

© Springer-Verlag 1977

Authors and Affiliations

  • A. Peter Czernilofsky
    • 1
  • Ekkehard Collatz
    • 1
  • Axel M. Gressner
    • 1
  • Ira G. Wool
    • 1
  • Ernst Küchler
    • 2
  1. 1.Department of BiochemistryUniversity of ChicagoChicagoUSA
  2. 2.Institut für Biochemie der Universität WienWien 9Austria

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