Cooperative binding of zinc to an aminoacyl-tRNA synthetase

Zinc binds to tetrameric alanyl-tRNA synthetase from Escherichia coli with a stoichiometry of one g-atom of zinc per enzyme subunit. The nature of this metal-protein interaction is investigated here through a series of equilibrium dialysis and intrinsic fluorescence experiments. The dialysis data show that zinc binds to this synthetase in a cooperative manner, with half-maximal zinc binding at 0.97 microM free zinc and a Hill coefficient of 1.9. The cooperative feature is also observed in the zinc-induced quenching of the protein intrinsic fluorescence, indicating that zinc binding induces a conformational change. This is the first report of cooperative binding of zinc to an aminoacyl-tRNA synthetase, and the data provide a rationale for the oligomeric structure of the synthetase specific for alanine.