PMID: 44203Jan 1, 1979

The twenty aminoacyl-tRNA synthetases from Escherichia coli. General separation procedure, and comparison of the influence of pH and divalent cations on their catalytic activities

D Kern, Jacques Lapointe


A general separation procedure of the twenty E. coli aminoacyl-tRNA synthetases including either a 105 000 g centrifugation or a poly-ethyleneglycol-dextran two-phases partition fractionation, and chromatographies on DEAE-cellulose, phosphocellulose and hydroxyapatite is described. The specific activities of the synthetases have been determined after each chromatographic step and compared to their respective activities in the 105 000 g supernatant. Some aminoacyl-tRNA synthetases were obtained at 80 per cent purity. The presence of phenylmethylsulfonyl fluoride does not significantly modify either the elution patterns of the synthetases during the various chromatographic steps or their specific activities. Thus, contrarily to enzymes from various eukaryotic organisms no significant inactivation of the E. coli aminoacyl-tRNA synthetases occurs via proteolytic processes during the purification procedure. The effects of various factors: pH, magnesium, and other bivalent cations including spermidine, were tested on the aminoacylation and the [32P] PPi-ATP isotope-exchange reactions, and the optimal aminoacylation and isotope-exchange conditions determined for 18 of the 20 E. coli aminoacyl-tRNA synthetases.


Mar 9, 2004·Journal of Molecular Biology·Valérie CampanacciChristian Cambillau
Jul 24, 2012·Nucleic Acids Research·Chih-Chi LiaoChien-Chia Wang
May 30, 2009·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Marc BaillyD Kern
Aug 3, 1999·Biological Chemistry·W FreistJ G Arnez
Oct 3, 1983·European Journal of Biochemistry·M H RatinaudR Julien
Jun 1, 1987·European Journal of Biochemistry·B LorberR Giege

Related Concepts

Cations, Divalent
Alkalescens-Dispar Group
Magnesium Measurement
Enzymes, antithrombotic
MT-TA gene
Triplet Codon-amino Acid Adaptor Activity
Transfer RNA
Proteolytic Enzyme

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