Probing the influences of urea on the interaction of sinomenine with human serum albumin by steady-state fluorescence

The binding of sinomenine to human serum albumin (HSA) in aqueous solution in the absence and presence of urea has been studied by fluorescence and the three-dimensional (3D) fluorescence at pH 7.40. Subdomain IIA binding site of human serum albumin (HSA) was characterized by examining the change in HSA fluorescence. The quenching rate constants and binding constants were calculated in the absence and presence of the denaturant. The results point to a static quenching mechanism operating in the complexes. However, the binding ability of sinomenine to denatured HSA is weaker than that of sinomenine to native HSA. Denaturation of HSA in the presence of urea is almost complete at [urea] ⩾ 8.0 M. Upon unfolding, two fluorescence peaks were observed. One peak was assigned to the fluorescence of Trp-214 residue in a polar environment, and the other peak was assigned to the fluorescence of tyrosine residues. Compared to the free HSA, the HSA-sinomenine complex is more stable in the presence of urea.

► The fluorescence of native and denatured HSA was quenched by sinomenine at pH 7.4.
► Fluorescence spectrum indicated urea-induced unfolding of native HSA.
► Compared to HSA, HSA-sinomenine complex is more stable in the presence of urea.
► Sinomenine was harder to bind to the denatured HSA.