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Reaction of Hg2+ Insertion into Cysteine Pairs Within Bovine Insulin Crystals Followed via Raman Spectroscopy

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Abstract

Chemical modifications of protein crystals may be achieved via soaking of reactants from their precipitating solution, through the solvent channel, into the protein matrix. We describe a Raman microscopy approach to follow mercury insertion into cysteine pairs within protein single crystals, via soaking in an aqueous Hg2+ solution. The method has been developed using bovine insulin as the model system. Applying an efficient mercuration protocol, consisting of a first step of disulphide bridge TCEP-induced reduction within the crystal, followed by overnight reaction with a HgCl2 solution, we obtained Hg-derivative crystals. Raman spectra collected on these derivative crystals, kept in the mother liquor, reveal a characteristic Raman band at 335 cm−1, which has been assigned to a –S–Hg–S– bridge. The analysis provides Raman-based markers of mercury binding to cysteines, and thus of mercury intoxication.

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Authors and Affiliations

  1. Department of Chemical Sciences, University of Naples “Federico II”, Complesso Universitario di Monte Sant’Angelo, Via Cintia, 80126, Naples, Italy

    Marco Caterino, Antonello Merlino, Anna Balsamo, Irene Russo Krauss & Alessandro Vergara

  2. CNR Institute of Biostructures and Bioimages, Naples, Italy

    Antonello Merlino & Alessandro Vergara

  3. Department of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”, Naples, Italy

    Silvia Parisi

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  1. Marco Caterino
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  2. Antonello Merlino
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  3. Anna Balsamo
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  4. Irene Russo Krauss
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  5. Silvia Parisi
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  6. Alessandro Vergara
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Correspondence to Alessandro Vergara.

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Caterino, M., Merlino, A., Balsamo, A. et al. Reaction of Hg2+ Insertion into Cysteine Pairs Within Bovine Insulin Crystals Followed via Raman Spectroscopy. J Solution Chem 43, 135–143 (2014). https://doi.org/10.1007/s10953-013-0066-y

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