Application of capillary zone electrophoresis to the study of interactions betweenBacillus subtilis tryptophanyl-tRNA synthetase and tRNATrp

Abstract

SummaryThe application of capillary zone electrophoresis to the study of interactions between Bacillus subtilis tryptophanyl-tRNA synthetase (TrpRS) and tRNATrp is described. Significant changes in peak shape of tRNATrp incubated with TrpRS indicated the occurrence of interactions between TrpRS and tRNATrp in pH 8.0 Tris-HC1 buffer containing 0.1 mmol L -1 EDTA and 1 mmol L -1 -5 mmol L -1 MgC12. Addition of Mg 2 § decreased the electrophoretic mobility of tRNATrp, which illustrated that conformation of tRNATrp depended on Mg 2+. The dissociation constant of the TrpRS-tRNATrp complex was estimated to be 0.63 lamol L -1 at 25 ~ in buffer solution.

Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2

“…However, the interaction between receptor and ligand lengthens the receptor (or ligand) zone and causes the peak broadening. The more ligand (or receptor) the sample contains, the wider the receptor (or ligand) peak becomes, and the lower the receptor (or ligand) peak height changes [38,39]. The changes are successively and regularly, which mean that it is not occasional or dues to other effects, but the interaction occurrence.…”

“…Based on the CZE/FA method, the peak height changes of receptor (or ligand) resulting from interacting with ligand (or receptor) can be used to estimate the binding constant [40,41]. Previous reports have successfully estimated the binding constants of two fast binding kinetic systems, TrpRS-tRNA [39], DNA-netropsin [42]. From this study, we demonstrated that fast on-and-off kinetic interactions could also be studied by CZE, but not only by ACE.…”

Recent advances in the study of biomolecular interactions by capillary electrophoresis

“…Weak binding is typically analyzed by ACE or FA [14]. According to recent observations [15,16], CZE can be also applied to study weak binding systems, in contrast to some former reports [11,17]. It was found in CZE that the peak height can also be used to evaluate the concentration of free ligand or receptor as in FA.…”

“…It was found in CZE that the peak height can also be used to evaluate the concentration of free ligand or receptor as in FA. When only small amounts of samples are available, CZE is a more appropriate mode to conduct weak binding assays [15,16]. In fact, CZE and FA are identical except for a difference in the length of the analyte zone, which means that in CZE, the receptor and ligand can be injected immediately after mixing, or after certain time of incubation.…”

“…Using CE, Zhang et al [16] studied the interaction between tryptophanyl-tRNA synthetase and trp tRNA of Bacillus subtilis with a fast and on-and-off kinetic interaction. The receptor-ligand association broadens the migration zones of the analytes.…”