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Fourth derivative UV-spectroscopy of proteins under high pressure II. Application to reversible structural changes

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Abstract

The structural basis and the thermodynamics of pressure induced reversible spectral transitions in the fourth derivative ultraviolet absorbance spectra of proteins were analysed as described in the preceding paper. Three proteins were studied: adrenodoxin (a small iron-sulphur protein that serves as an electron donor for cytochrome P450scc), ribonuclease A, and methanol dehydrogenase (a tetrameric protein). Fourth derivative spectroscopy is used to probe important mechanistic aspects of these proteins. For adrenodoxin, the results suggest that one or two phenylalanines interact with the iron-sulphur redox centre. High pressure denaturation of ribonuclease leads to a molten globule like structure that also occurs as an intermediate in the high temperature induced denaturation process. This state is characterised by the local dielectric constant in the vicinity of tyrosines. Methanol dehydrogenase was found to be very stable towards pressure. High pressure appears to strengthen the interaction between the two α-subunits possibly through the increased interaction of four tryptophans with other aromatic amino acids.

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Author notes

  1. V. V. Mozhaev

    Present address: Chemistry Department, Moscow State University, 000958, Moscow, Russia

Authors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale, U128, BP 5051, F-34033, Montpellier cedex, France

    R. Lange, N. Bec & V. V. Mozhaev

  2. Kluyver Laboratory for Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC, Delft, The Netherlands

    J. Frank

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Lange, R., Bec, N., Mozhaev, V.V. et al. Fourth derivative UV-spectroscopy of proteins under high pressure II. Application to reversible structural changes. Eur Biophys J 24, 284–292 (1996). https://doi.org/10.1007/BF00180369

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  • DOI: https://doi.org/10.1007/BF00180369

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