Yeast ribosomal protein L26 is located at the ribosomal subunit interface as determined by chemical cross-linking

J. Villarreal, J. C. Lee

Research output: Contribution to journalArticle

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Abstract

Previous studies suggested that yeast ribosomal protein L26 was a candidate for the ribosomal subunit interface region. The present study used protein-protein cross-linking to identify neighboring proteins in intact 80S ribosomes of Saccharomyces cerevisiae. To facilitate identification of cross-linked pairs involving L26, 80S ribosomes were first treated with 5-iodoacetamidofluorescein to selectively label L26. Protein cross-links were produced with dithiobis[succinimidyl] propionate or N-succinimidyl-3-[2-pyridyldithio] propionate and analyzed by electrophoresis on two-dimensional diagonal polyacrylamide gels containing SDS. L26 was detected under UV and its crosslinked partner, detectable after staining with Coomassie blue, was located below the diagonal and was coincident with L26 on a single vertical axis. The identity of the partner was determined by its co-migration with 60S and 40S ribosomal protein markers on two-dimensional gels. Two protein pairs involving L26 and 40S subunit proteins were identified. The finding provided experimental evidence to support that L26 is located at the ribosomal subunit interface. A model that incorporates the present findings and the published data on intra-subunit protein pairs is proposed for the yeast 80S ribosome.

Original languageEnglish (US)
Pages (from-to)321-324
Number of pages4
JournalBiochimie
Volume80
Issue number4
DOIs
StatePublished - Apr 1998

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Keywords

  • Chemical cross-linking
  • Ribosome
  • Ribosome subunit interface
  • Yeast

ASJC Scopus subject areas

  • Biochemistry

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