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Steady-state and femtosecond photoinduced processes of blepharismins bound to alpha-crystallin

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Abstract

The interaction of blepharismin (BP) and oxyblepharismin (OxyBP) with bovine alpha-crystallin (BAC) has been studied both by steady-state and femtosecond spectroscopy, with the aim of assessing the possible phototoxicity of these compounds toward the eye tissues. We showed that these pigments form with BAC potentially harmful ground-state complexes, the dissociation constants of which have been estimated to be 6 ± 2 µmol L−1 for OxyBP and 9 ± 4 μmol L−1 for BP. Irradiation with steady-state visible light of solutions of blepharismins in the presence of BAC proved to induce a quenching of both the pigment and the intrinsic protein fluorescences. These effects were tentatively rationalized in terms of structural changes of alpha-crystallin. On the other hand, femtosecond transient absorption spectroscopy was used to check the occurrence of any type I photoactivity of oxyblepharismin bound to alpha-crystallin. The existence of a particular type of fast photoinduced reaction, not observed in former studies with human serum albumin but present in the natural oxyblepharismin-binding protein, could here be evidenced but no specific reaction was observed during the first few nanoseconds after excitation. Partial denaturation of alpha-crystallin was however found to alter the excited-state behaviour of its complex with oxyblepharismin, making it partly resemble that of free oxyblepharismin in solution.

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Authors and Affiliations

  1. National Institute of Laser Enhanced Sciences (NILES), Cairo University, Giza, Egypt

    T. Youssef

  2. Département de Chimie UMR 8640-PASTEUR, Ecole Normale Supérieure, 24 rue Lhomond, 75005, Paris, France

    J. Brazard, C. Ley, F. Lacombat, P. Plaza & M. M. Martin

  3. CNR Istituto di Biofisica, Via G. Moruzzi 1, 56100, Pisa, Italy

    A. Sgarbossa, G. Checcucci & F. Lenci

Authors
  1. T. Youssef
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  2. J. Brazard
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  3. C. Ley
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  4. F. Lacombat
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  5. P. Plaza
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  6. M. M. Martin
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  7. A. Sgarbossa
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Correspondence to P. Plaza or A. Sgarbossa.

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Youssef, T., Brazard, J., Ley, C. et al. Steady-state and femtosecond photoinduced processes of blepharismins bound to alpha-crystallin. Photochem Photobiol Sci 7, 844–853 (2008). https://doi.org/10.1039/b800848e

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