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Isolation and characterization of a tRNA(guanine-7-)-methyltransferase from Salmonella typhimurium

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Summary

The tRNA modifying enzyme, S-adenosylmethionine:tRNA(guanine-7-)-methyltransferase, has been extensively purified from Salmonella typhimurium. A rapid and efficient purification method using phosphocellulose chromatography followed by ammonium sulfate precipitation and Sephadex G-100 gel filtration is described. The enzyme appears to be a single polypeptide chain with a molecular weight of approximately 25 000–30 000 daltons. The Km for S-adenosylmethionine and for undermethylated tRNA is 53 μ M and 3.4 μM, respectively. The methylation reaction is dependent on added monovalent or divalent cations; 5 mM spermidine, 3 mM MgCl2 and 1 mM spermine are the most effective. The enzyme, though not homogeneous, is free from contaminating ribonucleases and other tRNA methyltransferases.

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Abbreviations

A260 unit:

the quantity of material contained in 1 ml of solution which has an absorbance of 1 at 260 nm when measured in a 1 cm lightpath cell

AdoMet:

S-adenosylmethionine

TCA:

trichloroacetic acid

SDS:

sodium dodecyl sulfate

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Author notes

  1. Gennaro Ciliberto

    Present address: European Molecular Biology Laboratory, Postfach 102209, 6900, Heidelberg, West Germany

Authors and Affiliations

  1. Istituto di Chimica Biologica, II Facoltà di Medicina e Chirurgia, Università di Napoli, Via Sergio Pansini 5, 80131, Napoli, Italy

    Alfredo Colonna, Gennaro Ciliberto, Rita Santamaria, Filiberto Cimino & Francesco Salvatore

Authors
  1. Alfredo Colonna
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  2. Gennaro Ciliberto
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  3. Rita Santamaria
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  4. Filiberto Cimino
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  5. Francesco Salvatore
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Colonna, A., Ciliberto, G., Santamaria, R. et al. Isolation and characterization of a tRNA(guanine-7-)-methyltransferase from Salmonella typhimurium . Mol Cell Biochem 52, 97–106 (1983). https://doi.org/10.1007/BF00224919

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  • DOI: https://doi.org/10.1007/BF00224919

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