Brief Communication | Published:

Metallo-β-lactamases withstand low Zn(II) conditions by tuning metal-ligand interactions

Nature Chemical Biology volume 8, pages 698700 (2012) | Download Citation

Abstract

A number of multiresistant bacterial pathogens inactivate antibiotics by producing Zn(II)-dependent β-lactamases. We show that metal uptake leading to an active dinuclear enzyme in the periplasmic space of Gram-negative bacteria is ensured by a cysteine residue, an unusual metal ligand in oxidizing environments. Kinetic, structural and affinity data show that such Zn(II)-cysteine interaction is an adaptive trait that tunes the metal binding affinity, thus enabling antibiotic resistance at restrictive Zn(II) concentrations.

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References

  1. 1.

    , & Acc. Chem. Res. 39, 721–728 (2006).

  2. 2.

    , & Chem. Rev. 105, 395–424 (2005).

  3. 3.

    et al. Biochemistry 37, 12404–12411 (1998).

  4. 4.

    , , , & Biochemistry 37, 10173–10180 (1998).

  5. 5.

    , , & J. Am. Chem. Soc. 130, 14207–14216 (2008).

  6. 6.

    et al. J. Am. Chem. Soc. 131, 10753–10762 (2009).

  7. 7.

    , & J. Am. Chem. Soc. 130, 15842–15851 (2008).

  8. 8.

    et al. J. Biol. Chem. 277, 24142–24147 (2002).

  9. 9.

    et al. J. Am. Chem. Soc. 130, 15852–15863 (2008).

  10. 10.

    & Biochemistry 45, 11012–11020 (2006).

  11. 11.

    et al. EMBO J. 14, 4914–4921 (1995).

  12. 12.

    et al. J. Mol. Biol. 357, 890–903 (2006).

  13. 13.

    , & Biochemistry 49, 7930–7938 (2010).

  14. 14.

    & Nat. Chem. Biol. 4, 148–151 (2008).

  15. 15.

    , , & FEBS Lett. 503, 1–6 (2001).

  16. 16.

    et al. Antimicrob. Agents Chemother. 44, 2304–2309 (2000).

  17. 17.

    , , , & J. Biol. Chem. 282, 30586–30595 (2007).

  18. 18.

    & J. Biol. Chem. 279, 26046–26051 (2004).

  19. 19.

    et al. J. Biol. Chem. 284, 28164–28171 (2009).

  20. 20.

    et al. J. Mol. Biol. 392, 1278–1291 (2009).

  21. 21.

    & Science 292, 2488–2492 (2001).

  22. 22.

    , & Chem. Rev. 109, 4644–4681 (2009).

  23. 23.

    , & Antimicrob. Agents Chemother. 53, 2908–2917 (2009).

Download references

Acknowledgements

This work has been supported by CONICET and by grants from the Howard Hughes Medical Institute, Agencia Nacional de Promoción Científica y Tecnológica, US National Institutes of Health (1R01AI100560 to A.J.V.) and Laboratório Nacional de Luz Síncrotron, Campinas, Brazil. A.J.V. is a fellow of the John Simon Guggenheim Foundation.

Author information

Author notes

    • Javier M González
    •  & Francisco J Medrano Martín

    Present addresses: Department of Pharmaceutical Sciences, University of Maryland at Baltimore, Baltimore, Maryland, USA (J.M.G.); University of Maryland Crystallography Shared Service, University of Maryland at Baltimore, Baltimore, Maryland, USA (J.M.G.) and Centro de Investigaciones Biológicas, Madrid, Spain (F.J.M.M.).

    • Javier M González
    •  & María-Rocío Meini

    These authors contributed equally to this work.

Affiliations

  1. Instituto de Biología Molecular y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.

    • Javier M González
    • , María-Rocío Meini
    • , Pablo E Tomatis
    • , Julia A Cricco
    •  & Alejandro J Vila
  2. Laboratório Nacional de Luz Síncrotron, Sao Paulo, Brazil.

    • Francisco J Medrano Martín

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Contributions

J.M.G., J.A.C., M.-R.M., P.E.T. and A.J.V. designed experiments and analyzed results. J.M.G., M.-R.M., P.E.T. and A.J.V. wrote the manuscript. J.M.G., J.A.C. and M.-R.M. expressed and purified proteins. J.M.G. and J.A.C. determined kinetic parameters and performed cobalt substitution. J.M.G. performed stopped-flow measurements. M.-R.M. determined the dissociation constants for Zn(II) by competition experiments and the activity dependence on Zn(II) concentration. J.M.G. and F.J.M.M. determined the crystal structures. J.A.C. designed and made plasmid constructs, determined minimum inhibitory concentrations and performed in vivo antibiotic sensitivity tests. J.A.C. and M.-R.M. performed periplasmic extracts and western blot assays.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Julia A Cricco or Alejandro J Vila.

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DOI

https://doi.org/10.1038/nchembio.1005

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