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


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).


Electrophoresis Polyacrylamide Dodecyl Sodium Dodecyl Sodium Dodecyl Sulfate 
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p-nitrophenoxycarbonyl-3H-phenylalanyl-tRNA yeast Phe (phenylalanine specific, isolated from yeast)


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




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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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