Glutathione complex formation with mercury(II) in aqueous solution at physiological pH.

The mercury(II) complexes formed in neutral aqueous solution with glutathione (GSH, here denoted AH(3) in its triprotonated form) were studied using Hg L(III)-edge extended X-ray absorption fine structure (EXAFS) and (199)Hg NMR spectroscopy, complemented with electrospray ionization mass spectrometric (ESI-MS) analyses. The [Hg(AH)(2)](2-) complex, with the Hg-S bond distances at 2.325 ± 0.01 Å in linear S-Hg-S coordination, and the (199)Hg NMR chemical shift at -984 ppm, dominates except at high excess of glutathione. In a series of solutions with C(Hg(II)) ∼17 mM and GSH/Hg(II) mole ratios rising from 2.4 to 11.8, the gradually increasing mean Hg-S bond distance corresponds to an increasing amount of the [Hg(AH)(3)](4-) complex. ESI-MS peaks appear at -m/z values of 1208 and 1230 corresponding to the [Na(4)Hg(AH)(2)(A)](-) and [Na(5)Hg(AH)(A)(2)](-) species, respectively. In another series of solutions at pH 7.0 with C(Hg(II)) ∼50 mM and GSH/Hg(II) ratios from 2.0 to 10.0, the Hg L(III)-edge EXAFS and (199)Hg NMR spectra show that at high excess of glutathione (∼0.35 M) about ∼70% of the total mercury(II) concentration is present as the [Hg(AH)(3)](4-) complex, with the average Hg-S bond distance 2.42 ± 0.02 Å in trigonal HgS(3) coordination. The proportions of HgS(n) species, n = 2, 3, and 4, quantified by fitting linear combinations of model EXAFS oscillations to the experimental EXAFS data in our present and previous studies were used to obtain stability constants for the [Hg(AH)(3)](4-) complex and also for the [Hg(A)(4)](10-) complex that is present at high pH. For Hg(II) in low concentration at physiological conditions (pH 7.4, C(GSH) = 2.2 mM), the relative amounts of the HgS(2) species [Hg(AH)(2)](2-), [Hg(AH)(A)](3-), and the HgS(3) complex [Hg(AH)(3)](4-) were calculated to be 95:2:3. Our results are not consistent with the formation of dimeric Hg(II)-GSH complexes proposed in a recent EXAFS study.