A mass spectrometric investigation of the binding of gold antiarthritic agents and the metabolite [Au(CN)2]- to human serum albumin.

Electrospray ionisation (ESI) mass spectrometry was used to examine the reactions of the clinically used antiarthritic agent [Au(S2O3)2]3-, and AuPEt3Cl, a derivative of another clinically used agent auranofin, with human serum albumin (HSA) obtained from a human volunteer. Both compounds reacted readily with HSA to form complexes containing one or more covalently attached gold fragments. In the case of AuPEt3Cl, binding was accompanied by the loss of the chloride ligand, while for [Au(S2O3)2]3- the mass spectral data indicated binding of Au(S2O3) groups. Experiments performed using HSA with Cys34 blocked by reaction with iodoacetamide were consistent with reaction of both gold compounds with this amino acid. Separate blocking experiments using diethylpyrocarbonate and AuPEt3Cl also provided evidence for histidine residues acting as lower-affinity binding sites for this gold compound. ESI mass spectra of solutions containing [Au(S2O3)2]3- or [Au(CN)2]-, and HSA, provided evidence for the formation of protein complexes in which intact gold molecules were non-covalently bound. In the case of [Au(S2O3)2]3-, these non-covalent complexes proved to be transitory in nature. However, for [Au(CN)2]- a non-covalent complex containing a single gold molecule bound to HSA was found to be stable, and constituted the main adduct formed in solutions containing low-to-medium Au-to-HSA ratios. Evidence was also obtained for the formation of a covalent adduct in which a single Au(CN) moiety was bonded to Cys34 of the protein. AuPEt3Cl reacted to a much lower extent with HSA that had Cys34 modified by formation of a disulfide bond to added cysteine, than with unmodified HSA. This suggests that the extent of modification of the protein in vivo may have an important influence on the transport and bioavailability of gold antiarthritic drugs.