engleski

Stability of the complex between yeast seryl- tRNA synthetase and tRNASer under different electrophoretic conditions

Noncovalent interactions of yeast homodimeric seryl-tRNA synthetase (SerRS) and cognate tRNASer were studied by the gel mobility shift assay and zone-interference gel electrophoresis performed under the same binding and electrophoretic conditions. Purified tRNASer as well as total yeast tRNA were applied as ligands. In the absence of Mg2+, SerRS: (tRNASer)1 noncovalent complex was detected only by zone-interference gel electrophoresis. Kd values determined in the presence and absence of Mg2+ were in the same range, suggesting that Mg2+ ions mainly influence dissociation-association kinetics of the complex, with minor contribution to its thermodynamic stability. Comparison of these two assays was shown to be useful in the analysis of thermodynamic and kinetic properties of protein:nucleic acids complexes.

aminoacyl-tRNA synthetase ; gel mobility shift assay ; zone-interference gel electrophoresis ; SerRS:tRNASer noncovalent complexes ; Mg2+ influence

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