Suppression of high-affinity ligand binding to the major glutathione S-transferase from Galleria mellonella by physiological concentrations of glutathione.

The major glutathione S-transferase from larvae of Galleria mellonella binds a number of synthetic triphenylmethane dyes with dissociation constants of the order of 10(-6) M or less. The organ distribution of the enzyme activity does not parallel the uptake of such dyes by the insect's organs in vivo. The affinity of the protein for such dyes is decreased by about an order of magnitude by the presence of glutathione in normal physiological concentration. This appears to be the cause of this protein's lack of efficacy as a 'ligandin' in vivo. The dyes appear to be acting as ineffective substrate analogues, binding at the catalytic site and impeding, in a reciprocal fashion, the binding of glutathione. Fluorescence-quenching titration and kinetic experiments together indicate the existence of a single ligand-binding and catalytic site per dimeric enzyme molecule.