Journal of Protein Chemistry

, Volume 19, Issue 6, pp 431–439 | Cite as

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

  • Pierre M. Viallet
  • Tuan Vo-Dinh
  • Anne-Cécile Ribou
  • Jean Vigo
  • Jean-Marie Salmon


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.

Fluorescence spectra fluorescence quenching energy transfer protein folding conformational changes structural biology tryptophan 


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Copyright information

© Plenum Publishing Corporation 2000

Authors and Affiliations

  • Pierre M. Viallet
    • 1
    • 2
  • Tuan Vo-Dinh
    • 3
  • Anne-Cécile Ribou
    • 1
  • Jean Vigo
    • 1
  • Jean-Marie Salmon
    • 1
  1. 1.Quantitative Microfluorometry Group, Laboratory of Physicochemical Biology of Integrated SystemsUniversity of PerpignanPerpignanFrance
  2. 2.Advanced Monitoring Development Group, Life Sciences DivisionOak Ridge National LaboratoryOak Ridge
  3. 3.Advanced Monitoring Development Group, Life Sciences DivisionOak Ridge National LaboratoryOak Ridge

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