The formation of Ni(II) complexes of GSH in conditions of 4-fold GSH excess over Ni(II) was studied by potentiometric titrations, UV-vis and CD spectroscopies, and magnetic susceptibility measurements. The following set of complexes was obtained in the pH range of 6-12: NiHL, Ni(2)L(2)(2)(-), NiHL(2)(3)(-), NiL(2)(4)(-), and NiH(-)(1)L(2)(5)(-). The first of these is an octahedral species, coordinated through the donors of the Glu moiety of GSH, while the remaining ones are largely square-planar, with participation of the thiol in Ni(II) coordination. Magnetic moments indicate the presence of a spin equilibrium for Ni(2)L(2)(2)(-), NiHL(2)(3)(-), and NiL(2)(4)(-) complexes. Phosphate ions apparently decompose the Ni(2)L(2)(2)(-) complex, converting it into a monomeric, high spin, ternary species. Among the molecular forms of GSH, HL(2)(-) is the one most susceptible to air oxidation, due to a presence of ionic interactions between its protonated amine and deprotonated thiol moieties. The complexation of Ni(II) accelerates air oxidation of GSH in alkaline solutions by a factor of 4, but this effect is absent at neutral pH. The damage to plasmid DNA by H(2)O(2) is facilitated by Ni(II) ions and inhibited by excess of GSH. However, the analysis of the concentration profile of this process indicates that octahedral Ni(II) complexes with GSH are involved in the formation of double strand breaks. Finally, numerical simulations of intracellular Ni(II) distribution, made possible by the determination of stability constants of Ni(II) complexes of GSH, indicate that histidine and ATP, rather than GSH, may act as ligands for Ni(II) in vivo. Altogether, our results suggest that the direct impact of GSH on Ni(II) toxicity may be of a limited character.