Half molecules of serine-specific transfer ribonucleic acids from yeast |
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Authors: | F Fittler H G Zachau |
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Affiliation: | Institut für Physiologische Chemie und Physikalische Biochemie der Universität München, German Federal Republic |
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Abstract: | The preparation and analysis of half molecules from tRNASer are described. Two pG-halves were isolated which differed only in the presence or absence of an acetyl group on the cytidylic acid residue at position 12. The CCA-half derived from tRNA1Ser was isolated pure, while the CCA-half derived from tRNA2Ser was isolated as a mixture with the CCA-half from tRNA1Ser from which the terminal CpCpA had been cleaved off.The acceptor activity of the combined complementary half molecules was 90% of the one of intact tRNASer. The Michaelis constant and maximal velocity of amino-acylation were found to be identical for tRNASer and the combined fragments.When half molecules were present at different ratios in aminoacylation studies it was found that one pG-half molecule can mediate the charging of several CCA-half molecules. There are indications that the CCA-half molecule alone can accept some serine. The CCA-half molecule alone can be aminoacylated to a rather high degree in the presence of an excess of tRNAoxSer or tRNASer-a and to a small degree in the presence of tRNAoxAla (yeast) but not at all in the presence of tRNAoxPhe or tRNAoxVal (E. coli).Combinations of half molecules from tRNASer with the opposite half molecules from tRNAPhe could not be aminoacylated with Ser or Phe or 15 other amino acids although one of the combinations was well associated according to gel electrophoresis and differential melting curves. |
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