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Analysis by two-dimensional polyacrylamide gel electrophoresis of the in vivo phosphorylation of ribosomal proteins derived from free and membrane-bound polysomes

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Abstract

Analysis of in vivo phosphorylation of mouse liver ribosomal proteins was performed by two-dimensional polyacrylamide gel electrophoresis following 32P-injection. Our method is special and differs from other eukaryotic systems reported in that all proteins separated on the first dimension gel are completely solubilized, moving quantitatively to the second dimension gel. Only ribosomes from polysomes were used, ensuring analysis of ribosomes actively engaged in protein synthesis. We resolved sixty-five distinct proteins from ribosomes from membrane bound or free polysomes. In both cases radioautography revealed similar labeled patterns with one highly phosphorylated ribosomal protein and five marginally labeled spots.

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Author notes

  1. Wayne L. Hoffman

    Present address: Dept. of Biophysics, The University of Texas, Southwestern Medical School, 75235, Dallas, Tex., USA

Authors and Affiliations

  1. Dept. of Anatomy and Developmental Biology Center, School of Medicine, Case Western Reserve University, 44106, Cleveland, Ohio, USA

    Wayne L. Hoffman & Joseph Ilan

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  1. Wayne L. Hoffman
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  2. Joseph Ilan
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Hoffman, W.L., Ilan, J. Analysis by two-dimensional polyacrylamide gel electrophoresis of the in vivo phosphorylation of ribosomal proteins derived from free and membrane-bound polysomes. Molecular Biology Reports 2, 219–224 (1975). https://doi.org/10.1007/BF00356991

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  • DOI: https://doi.org/10.1007/BF00356991

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