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Protein structural changes induced by glutathione-coated CdS quantum dots as revealed by Trp phosphorescence

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Abstract

We evaluated the potential of tryptophan (Trp) phosphorescence spectroscopy for investigating conformational states of proteins involved in interaction with nanoparticles. Characterization of protein–nanoparticle interaction is crucial in assessing biological hazards related to use of nanoparticles. We synthesized glutathione-coated CdS quantum dots (GSH-CdS), which exhibited an absorption peak at 366 nm, indicative of 2.4 nm core size. Chemical analysis of purified GSH-CdS suggested an average molecular formula of GSH18S56Cd60. Investigations were conducted on model proteins varying in terms of isoelectric point, degree of burial of the Trp probe, and quaternary structure. GSH-CdS fluorescence measurements showed improvement in nanoparticle quantum yield induced by protein interaction. Trp phosphorescence was used to examine the possible perturbations in the protein native fold induced by GSH-CdS. Phosphorescence lifetime measurements highlighted significant conformational changes in some proteins. Despite their small size, GSH-CdS appeared to interact with more than one protein molecule. Rough determination of the affinity of GSH-CdS for proteins was derived from the change in phosphorescence lifetime at increasing nanoparticle concentrations. The estimated affinities were comparable to those observed for specific protein–ligand interactions and suggest that protein–nanoparticle interaction may have a biological impact.

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Acknowledgments

The technical assistance of Alessandro Puntoni is duly acknowledged.

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Authors and Affiliations

  1. Institute of Biophysics, via G. Moruzzi, 1, 56124, Pisa, Italy

    E. Gabellieri, P. Cioni, E. Balestreri & E. Morelli

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  1. E. Gabellieri
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  2. P. Cioni
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  3. E. Balestreri
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  4. E. Morelli
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Correspondence to E. Gabellieri.

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Special Issue: SIBPA 2011 Meeting.

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Gabellieri, E., Cioni, P., Balestreri, E. et al. Protein structural changes induced by glutathione-coated CdS quantum dots as revealed by Trp phosphorescence. Eur Biophys J 40, 1237–1245 (2011). https://doi.org/10.1007/s00249-011-0736-x

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  • DOI: https://doi.org/10.1007/s00249-011-0736-x

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