Native fluorescence and mag-indo-1-protein interaction as tools for probing unfolding and refolding sequences of the bovine serum albumin subdomain in the presence of guanidine hydrochloride

• Published in: Journal of Protein Chemistry 2000-08, Vol.19 (6), p.431-439
• Main Authors: Viallet, P M, Vo-Dinh, T, Ribou, A C, Vigo, J, Salmon, J M
• Format: Article
• Language: English
• Subjects: Animals; Biophysics; Bovine serum albumin; Calcium - chemistry; Cattle; Energy transfer; Fluorescence; Fluorescent Dyes - chemistry; Guanidine - pharmacology; Guanidine hydrochloride; Human serum albumin; Humans; Indo-1; Indoles - chemistry; Models, Molecular; Molecular biology; Protein Binding - drug effects; Protein Conformation - drug effects; Protein Denaturation; Protein Folding; Proteins; Serum albumin; Serum Albumin, Bovine - chemistry; Spectrometry, Fluorescence; Tryptophan; Tryptophan - metabolism
• ISSN: 0277-8033; 1572-3887; 1573-4943
• DOI: 10.1023/a:1026589012724

Changes in the fluorescence spectrum of tryptophans Trp 134 and Trp 212 in bovine serum albumin (BSA) and of Trp 214 of human serum albumin in the presence of the chaotropic agent guanidine hydrochloride (Gnd) were studied. A detailed analysis of the fluorescence spectrum of native BSA yielded the fluorescence spectrum for each tryptophan of BSA. Modifications in the binding of Mag-indo-1 to BSA, which results in a specific quenching of the fluorescence spectrum of Trp 134 associated with an energy transfer from Trp 134 to the protein-bound Mag-indo-1, were also investigated. Changes occurring when the Gnd concentration is decreased stepwise cover a larger concentration scale of Gnd than the reverse protocol, allowing one to suggest that the resulting conformational changes in the subdomain IA of BSA involve at least three different steps.