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Isolation and binding properties of leucyl-tRNA synthetase from Escherichia coli MRE 600

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Summary

A procedure for the large-scale isolation of leucyl-tRNA synthetase from E. coli MRE 600 is described: The enzyme was purified about 320-fold to homogeneity by precipitation with cetyl-trimethyl-ammonium bromide, two consecutive chromatographies on DEAE-cellulose and three on hydroxyapatite with an over-all yield of 4%.

The molecular weight of leucyl-tRNA synthetase from E. coli MRE 600 was found to be 99 000 daltons. Binding studies by ultracentrifugation and equilibrium partition showed that the enzyme binds leucine, leucyl-adenylate and tRNALeu, each in a 1 : 1 stoichiometry. For ATP only a very weak binding to the enzyme could be observed, which did not allow the evaluation of the complex stoichiometry. The presence of ATP was not required for the binding of leucine or tRNA to leucyl-tRNA synthetase from E. coli MRE 600.

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Authors and Affiliations

  1. Gesellschaft für Biotechnologische Forschung mbH., D-3300 Braunschweig-Stöckheim, Mascheroder Weg 1

    Stefica Granda, Helmut Hustedt, Josef Flossdorf & Maria-Regina Kula

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  1. Stefica Granda
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  2. Helmut Hustedt
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  3. Josef Flossdorf
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  4. Maria-Regina Kula
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Granda, S., Hustedt, H., Flossdorf, J. et al. Isolation and binding properties of leucyl-tRNA synthetase from Escherichia coli MRE 600. Mol Cell Biochem 24, 175–181 (1979). https://doi.org/10.1007/BF00220736

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  • DOI: https://doi.org/10.1007/BF00220736

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