Skip to main content
SpringerLink
Log in

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

  • Published:
Journal of Solution Chemistry Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Park, J.D., Zheng, W.: Human exposure and health effects of inorganic and elemental mercury. J. Prev. Med. Public Health 45, 344–352 (2012)

    Article  Google Scholar 

  2. Karagas, M.R., Choi, A.L., Oken, E., Horvat, M., Schoeny, R., Kamai, E., Cowell, W., Grandjean, P., Korrick, S.: Evidence on the human health effects of low-level methylmercury exposure. Environ. Health Perspect. 120, 799806 (2012)

    Article  Google Scholar 

  3. Aschner, M., Onishchenko, N., Ceccatelli, S.: Toxicology of alkylmercury compounds. Metal Ions Life Sci. 7, 403–434 (2010)

    Article  CAS  Google Scholar 

  4. Garcia, J.J., Martinez-Ballarin, E., Millan-Plano, S., Allue, J.L., Albendea, C., Fuentes, L., Escanero, J.F.: Effects of trace elements on membrane fluidity. J. Trace Elem. Med Biol. 19, 19–22 (2005)

    Article  CAS  Google Scholar 

  5. Houston, M.C.: Role of mercury toxicity in hypertension, cardiovascular disease, and stroke. J. Clin. Hypertens. (Greenwich) 13, 621–637 (2011)

    Article  CAS  Google Scholar 

  6. Koedrith, P., Seo, Y.R.: Advances in carcinogenic metal toxicity and potential molecular markers. Int. J. Mol. Sci. 12, 9576–9595 (2011)

    Article  CAS  Google Scholar 

  7. Valko, M., Morris, H., Cronin, M.T.: Metals, toxicity and oxidative stress. Curr. Med. Chem. 12, 1161–1208 (2005)

    Article  CAS  Google Scholar 

  8. Bernhoft, R.A.: Mercury toxicity and treatment: a review of the literature. J. Environ. Public Health. 2012, 460–508 (2012)

    Article  Google Scholar 

  9. Guzzi, G.La., Porta, C.A.: Molecular mechanisms triggered by mercury. Toxicology 244, 1–12 (2008)

    Article  CAS  Google Scholar 

  10. Vas, J., Monestier, M.: Immunology of mercury. Ann. N. Y. Acad. Sci. 1143, 240–267 (2008)

    Article  CAS  Google Scholar 

  11. Carvalho, C.M., Chew, E.H., Hashemy, S.I., Lu, J., Holmgren, A.: Inhibition of the human thioredoxin system. J. Biol. Chem. 283, 11913–11923 (2008)

    Article  CAS  Google Scholar 

  12. Brooks, P., Davidson, N.: Mercury(II) complexes of imidazole and histidine. J. Am. Chem. Soc. 82, 2118–2123 (1959)

    Article  Google Scholar 

  13. Shim, S.M., Dorworth, L., Lasrado, J., Santerre, C.: Mercury and fatty acids in canned tuna, salmon, and mackerel. J. Food Sci. 69, C681–C684 (2004)

    Article  CAS  Google Scholar 

  14. Zamora, F., Kunsman, M., Sabat, M., Lippert, B.: Metal-stabilized rare tautomers of nucleobases. 6. Imino tautomer of adenine in a mixed-nucleobase complex of mercury(II). Inorg. Chem. 36, 1583–1587 (1997)

    Article  CAS  Google Scholar 

  15. Corbeil, M.C., Beauchamp, A.L., Alex, S., Savoie, R.: Interaction of the methylmercury cation with glycine and alanine: a vibrational and X-ray diffraction study. Can. J. Chem. 64, 1876–1884 (1986)

    Article  CAS  Google Scholar 

  16. Saunders, C., Burford, N., Werner-Zwanzinger, U., McDonald, R.: Preparation and comprehensive characterization of [Hg6(alanine)4(NO3)4]·H2O. Inorg. Chem. 47, 3693–3699 (2008)

    Article  CAS  Google Scholar 

  17. Kalaiselvi, D., Kumar, R.M., Jayave, R.: Redetermination of poly[l-chloroheptachlorido-13-L-proline-12-L-proline tetramercury(II)]. Acta Crystallogr. Sect. E E64, m1048–m1049 (2008). Structure Reports

    Article  Google Scholar 

  18. Saunders, C.: Isolation of lead and mercury amino acid complexes with characterization in the solid state, the solution state, and gas state. PhD thesis, Department of Chemistry, Dalhouse University (2009)

  19. Taylor, N., Carty, A.: Nature of mercury(II) ion–l-cysteine complexes implicated in mercury biochemistry. J. Am. Chem. Soc. 99, 6143–6145 (1977)

    Article  CAS  Google Scholar 

  20. Adams, M.J., Hodgkin, D.C., Raeburn, U.A.: Crystal structure of a complex of mercury(II) chloride and histidine hydrochloride. J. Chem. Soc. A 2632–2635 (1970)

  21. Corbeil, M.C., Beauchamp, A.L.: Conformational changes in serine on methylmercury complexation. Can. J. Chem. 66, 1379–1385 (1988)

    Article  CAS  Google Scholar 

  22. Book, L., Carty, A.J., Chieh, C.: Five-coordinated halogenomercurate(II) complexes: crystal structures of bis(l-tryptophanium) trichloromercurate(II) and bis(d, l-homocysteine thiolactonium) tetrabromomercurate(II). Can. J. Chem. 59, 138–143 (1981)

    Article  CAS  Google Scholar 

  23. Alcock, N.W., Lampe, P.A., Moore, P.: Crystal structures of methyl(L-tyrosinato) mercury(II) monohydrate and [L-(2-amino-4-phenylbutanoato)] methylmercury(II). J. Chem. Soc. Dalton Trans. 1324–1328 (1978)

  24. Vergara, A., Merlino, A., Pizzo, E., D’Alessio, G., Mazzarella, L.: A novel method for detection of selenomethionine incorporation in protein crystals via Raman microscopy. Acta Crystallogr. Sect. D 64, 167–171 (2008)

    Article  CAS  Google Scholar 

  25. Carey, P.R., Dong, J.: Following ligand binding and ligand reactions in proteins via Raman crystallography. Biochemistry 43, 8885–8893 (2004)

    Article  CAS  Google Scholar 

  26. Vergara, A., Franzese, M., Merlino, A., Bonomi, G., Verde, C., Giordano, D., Mazzarella, L.: Correlation between hemichrome stability and the Root effect in tetrameric hemoglobins. Biophys. J. 97, 866–874 (2009)

    Article  CAS  Google Scholar 

  27. Zheng, R., Zheng, X., Dong, J., Carey, P.R.: Proteins can convert to β-sheet in single crystals. Protein Sci. 13, 1288–1294 (2004)

    Article  CAS  Google Scholar 

  28. Merlino, A., Verde, C., di Prisco, G., Mazzarella, L., Vergara, A.: Reduction of ferric hemoglobin from Trematomus bernacchii in a partial bis-histidyl state produces a deoxy coordination even when encapsulated into the crystal phase. J. Spectrosc. 22, 143–152 (2008)

    Article  CAS  Google Scholar 

  29. Carpentier, P., Royant, A., Weik, M., Bourgeois, D.: Advances in spectroscopic methods for biological crystals. 2. Raman spectroscopy. Appl. Cryst. 40, 1113–1122 (2007)

    Article  CAS  Google Scholar 

  30. Katona, G., Carpentier, P., Nivière, V., Amara, P., Adam, V., Ohana, J., Bourgeois, D.: Raman-assisted crystallography reveals end-on peroxide intermediates in a nonheme iron enzyme. Science 316, 449–453 (2007)

    Article  CAS  Google Scholar 

  31. Merlino, A., Fuchs, M.R., Pica, A., Balsamo, A., Dworkowski, F.S.N., Pompidor, G., Mazzarella, L., Vergara, A.: Selective X-ray-induced NO photodissociation in haemoglobin crystals: evidence from a Raman-assisted crystallographic study. Acta Crystallogr. Sect. D 69, 137–140 (2013)

    Article  CAS  Google Scholar 

  32. Vitagliano, L., Vergara, A., Bonomi, G., Merlino, A., Verde, C., di Prisco, G., Howes, B.D., Smulevich, G., Mazzarella, L.: Spectroscopic and crystallographic characterization of a tetrameric hemoglobin oxidation reveals structural features of the functional intermediate relaxed/tense state. J. Am. Chem. Soc. 130, 10527–10535 (2008)

    Article  CAS  Google Scholar 

  33. Vergara, A., D’Errico, G., Montesarchio, D., Mangiapia, G., Paduano, L., Merlino, A.: Interaction of anticancer ruthenium compounds with proteins: high resolution X-ray structures and Raman microscopy studies of the adduct between hen egg white lysozyme and AziRu. Inorg. Chem. 52, 4157–4159 (2013)

    Article  CAS  Google Scholar 

  34. Smulevich, G., Feis, A., Howes, B.D.: Fifteen years of Raman spectroscopy of engineered heme containing peroxidases: what have we learned? Acc. Chem. Res. 38, 433–440 (2005)

    Article  CAS  Google Scholar 

  35. Vergara, A., Vitagliano, L., Merlino, A., Sica, F., Marino, K., Verde, C., di Prisco, G., Mazzarella, L.: An order-disorder transition plays a role in switching off the Root effect in fish hemoglobins. J. Biol. Chem. 285, 32568–32575 (2010)

    Article  CAS  Google Scholar 

  36. Annunziata, O., Vergara, A., Paduano, L., Sartorio, R., Miller, D.G., Albright, J.G.: Precision of interferometric diffusion coefficients in a four-component system relevant to protein crystal growth: lysozyme-tetra(ethylene glycol)–NaCl–H2O. J. Phys. Chem. B 107, 6590–6597 (2003)

    Article  CAS  Google Scholar 

  37. Vergara, A., Annunziata, O., Paduano, L., Miller, D.G., Albright, J.G., Sartorio, R.: Multicomponent diffusion in crowded solutions. 2. Mutual diffusion in the ternary system tetra(ethylene glycol)–NaCl–water. J. Phys. Chem. B 108, 2764–2772 (2004)

    Article  CAS  Google Scholar 

  38. Annunziata, O., Vergara, A., Paduano, L., Sartorio, R., Miller, D.G., Albright, J.G.: Quaternary diffusion coefficients in a protein–polymer–salt–water system determined by Rayleigh interferometry. J. Phys. Chem. B 113, 13446–13453 (2009)

    Article  CAS  Google Scholar 

  39. Badger, J., Caspar, D.L.: Water structure in cubic insulin crystals. Proc. Natl. Acad. Sci. USA 88, 622–626 (1991)

    Article  CAS  Google Scholar 

  40. Smith, G.D., Pangborn, W.A., Blessing, R.H.: The structure of T6 bovine insulin. Acta Crystallogr. Sect. D 61, 1476–1482 (2005)

    Article  Google Scholar 

  41. Sperling, R., Steinberg, I.Z.: Simultaneous reduction and mercuration of disulfide bond A6-A11 of insulin by monovalent mercury. Biochemistry 13, 2007–2013 (1974)

    Article  CAS  Google Scholar 

  42. Dong, J., Swift, K., Matayoshi, E., Nienaber, V.L., Weitzberg, M., Rockway, T., Carey, P.R.: Probing inhibitors binding to human urokinase crystals by Raman microscopy: implications for compound screening. Biochemistry 40, 9751–9757 (2001)

    Article  CAS  Google Scholar 

  43. Zako, T., Sakono, M., Hashimoto, N., Ihara, M., Maeda, M.: Bovine insulin filaments induced by reducing disulfide bonds show a different morphology, secondary structure, and cell toxicity from intact insulin amyloid fibrils. Biophys. J. 96, 3331–3340 (2009)

    Article  CAS  Google Scholar 

  44. Zhu, G., Zhu, X., Fan, Q., Wan, X.: Raman spectra of amino acids and their aqueous solutions. Acta Spectrochim. A 78, 1187–1195 (2011)

    Article  Google Scholar 

  45. During, J.R., Cox, A.W.: Spectra and structure of organophosphorus compounds. XIV. Infrared and Raman spectra, vibrational assignment and the asymmetric potential function for ethylphosphine and ethylphosphine-d2. J. Chem. Phys. 63, 2303–2310 (1975)

    Article  Google Scholar 

  46. Jalilehvand, F., Leung, B.O., Izadifard, M., Damian, E.: Mercury(II) cysteine complexes in alkaline aqueous solution. Inorg. Chem. 45, 66–73 (2005)

    Article  Google Scholar 

  47. Hoffmann, G.G., Brockner, W., Steinfatt, I.: Bis(n-alkanethiolato) mercury(II) compounds, Hg(SC n H2n+1)2 (n = 1 to 10, 12): preparation methods, vibrational spectra, GC/MS investigations, and exchange reactions with diorganyl disulfides. Inorg. Chem. 40, 977–985 (2001)

    Article  CAS  Google Scholar 

  48. Fahey, R.C., Hunt, J.S., Windham, G.C.: On the cysteine and cystine content of proteins. Differences between intracellular and extracellular proteins. J. Mol. Evol. 10, 155–160 (1977)

    Article  CAS  Google Scholar 

  49. Hansen, R.E., Roth, D., Winther, J.R.: Quantifying the global cellular thiol-disulfide status. Proc. Natl. Acad. Sci. USA 106, 422–437 (2009)

    Article  CAS  Google Scholar 

Download references

Author information

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

Authors
  1. Marco Caterino
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Antonello Merlino
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anna Balsamo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Irene Russo Krauss
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Silvia Parisi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Alessandro Vergara
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alessandro Vergara.

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10953-013-0066-y

Keywords

Search

Navigation