Isolation of Threonyl–tRNA synthetase from Thermos thermophilus

Authors

  • A. D. Yaremchuk Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR Kiev, USSR Author
  • M. A. Tukalo Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR Kiev, USSR Author
  • S. P. Egorova Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR Kiev, USSR Author
  • G. Kh. Matsuka Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR Kiev, USSR Author

DOI:

https://doi.org/10.7124/bc.0000B0

Abstract

A method for isolation of high-purified threonyl-tRNA synthetase from T. thermophllus is described, including ammonium sulfate fractionation, chromatography on DEAE-sepharose, hydroxyapatite, heparine-sepharose and hydrophobic chromatography on Toyopearl HW-65. Yield of the purified enzyme was 9 mg from 1 kg of T. thermophllus cells. The enzyme is a dimer protein (a2 type) with molecular mass of 148 kDa.

References

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Hedrick JL, Smith AJ. Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis. Arch Biochem Biophys. 1968;126(1):155-64.

Weber K, Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969;244(16):4406-12.

Hennecke H, Bock A, Thomale J, Nass G. Threonyl-transfer ribonucleic acid synthetase from Escherichia coli: subunit structure and genetic analysis of the structural gene by means of a mutated enzyme and of a specialized transducing lambda bacteriophage. J Bacteriol. 1977;131(3):943-50.

Yamada H. Rapid purification of threonyl-tRNA synthetase from Saccharomyces carlsbergensis by affinity elution from phosphocellulose. J Biochem. 1978;83(6):1583-9.

Published

1989-03-20

Issue

Section

Short Communications