Identification of neighboring protein pairs in the 60 S ribosomal subunits from Saccharomyces cerevisiae by chemical cross-linking

R. H. Xiang, J. C. Lee

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Protein-protein cross-linking was used to determine the spatial arrangement of proteins within the 60 S ribosomal subunits of Saccharomyces cerevisiae. Protein cross-links were generated by treatment of intact ribosomal subunits with dimethyl 3,3'-dithiobispropionimidate. Proteins were extracted from the treated subunits and fractionated by Cm-cellulose chromatography. Cross-linked proteins in these fractions were analyzed by electrophoresis on two-dimensional diagonal polyacrylamide gels containing sodium dodecyl sulfate. Component members of cross-linked pairs were radiolabeled with 125I and identified by two-dimensional gel electrophoresis and comparison with nonradioactive ribosomal protein markers. Seventeen pairs involving 16 of the 45 60 S subunit proteins were identified. Several proteins were detected in numerous cross-linked dimers and were used as foci for constructing a model depicting the arrangement of proteins within the 60 S ribosomal subunit. The model also incorporated previously published data on structure and function of proteins from the yeast 60 S subunit.

Original languageEnglish (US)
Pages (from-to)10542-10546
Number of pages5
JournalJournal of Biological Chemistry
Volume264
Issue number18
StatePublished - 1989

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Ribosome Subunits
Yeast
Saccharomyces cerevisiae
Proteins
Electrophoresis
Fungal Proteins
Ribosomal Proteins
Protein S
Electrophoresis, Gel, Two-Dimensional
Protein Subunits
Cellulose
Sodium Dodecyl Sulfate
Chromatography
Dimers
Gels

ASJC Scopus subject areas

  • Biochemistry

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Identification of neighboring protein pairs in the 60 S ribosomal subunits from Saccharomyces cerevisiae by chemical cross-linking. / Xiang, R. H.; Lee, J. C.

In: Journal of Biological Chemistry, Vol. 264, No. 18, 1989, p. 10542-10546.

Research output: Contribution to journalArticle

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