Interaction of human serum albumin with oxovanadium ions studied by FT-IR spectroscopy and gel and capillary electrophoresis

Abstract

Some oxovanadium compounds have shown potential to inhibit RNase activity, while at the same time not inhibiting DNase activity. Some vanadyl complexes also inhibit protein synthesis in rabbit reticulocytes, but induce activation of protein–tyrosine kinase. To gain an insight into the interaction of oxovanadium ions with proteins, the present study was designed to examine the bindings of VOSO4and NaVO3salts with human serum albumin (HSA) in aqueous solution at physiological pH with metal ion concentrations of 0.0001 to 1 mM and HSA (fatty acid free) concentration of 2% w/v. Gel and capillary electrophoresis (CE) and Fourier transform infrared (FT-IR) spectroscopic methods were used to determine the metal ion binding mode, association constant, and the secondary structure of the protein in the presence of the oxovanadium compounds. Gel electrophoresis results showed that a maximum of 20 vanadyl cations (VO2+) are bound per HSA molecule with strong (K1= 7.0 × 107M–1) and weak (K2= 6.5 × 105M–1) bindings. Similarly, capillary electrophoresis showed two major bindings for vanadyl cation with K1= 1.2 × 108M–1and K2= 8.5 × 105M–1, whereas vanadate (VO–3) has only a weak binding affinity (K = 6.0 × 103M–1) with HSA molecule. The VO–3binds mainly to the lysine ε-amino NH+3groups, while VO2+binds possibly to the histidine nitrogen atom and the N-terminal of the α-amine residue. Infrared spectroscopic analysis showed metal ion binding results in major protein secondary structural changes from that of the α-helix (55.0 to 43–44%) to the β-sheet (22.0 to 23–26%), β-antiparallel (12.0 to 13–16%), and turn (11.0 to 17–18%), at high metal ion concentration. The observed spectral changes indicate a partial unfolding of the protein structure, in the presence of oxovanadium ions.Key words: oxovanadium, protein, binding mode, binding constant, secondary structure, electrophoresis, FT-IR spectroscopy.