Time-resolved fluorescence studies on bovine serum albumin denaturation process.
The denaturation of Bovine Serum Albumin (BSA) by a chaotropic agent, guanidinium hydrochloride (GuH+Cl-) was studied by fluorescence lifetime analysis. The BSA was labelled with 1-anilino-8-naphthalene sulfonate (ANS) at two different molar ratios (1:1) and (1:10). The non-exponential fluorescence kinetics of the BSA-ANS complex at different stages of denaturation is analysed using three different models: a discrete tri-exponential sum, stretched exponential, and Gaussian lifetime distribution. In all cases, the fluorescence decay times decreased with protein denaturation. The results from the models show that there are at least two different binding sites located in the BSA protein with different water accessibility.
Togashi
Denisio M
DM
Department of Chemistry and National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland. denisio.togashi@nuigalway.ie
Ryder
Alan G
AG
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry; D000818:Animals; D002417:Cattle; D005456:Fluorescent Dyes / Q000737:chemistry; D019791:Guanidine / Q000737:chemistry; D011489:Protein Denaturation; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013997:Time Factors
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D012710:Serum Albumin, Bovine; C027132:1-anilino-8-naphthalenesulfonate; D019791:Guanidine