Article | Published:

Nanomechanical detection of antibiotic–mucopeptide binding in a model for superbug drug resistance

Nature Nanotechnology volume 3, pages 691696 (2008) | Download Citation

Subjects

Abstract

The alarming growth of the antibiotic-resistant superbugs methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE) is driving the development of new technologies to investigate antibiotics and their modes of action. We report the label-free detection of vancomycin binding to bacterial cell wall precursor analogues (mucopeptides) on cantilever arrays, with 10 nM sensitivity and at clinically relevant concentrations in blood serum. Differential measurements have quantified binding constants for vancomycin-sensitive and vancomycin-resistant mucopeptide analogues. Moreover, by systematically modifying the mucopeptide density we gain new insights into the origin of surface stress. We propose that stress is a product of a local chemical binding factor and a geometrical factor describing the mechanical connectivity of regions activated by local binding in terms of a percolation process. Our findings place BioMEMS devices in a new class of percolative systems. The percolation concept will underpin the design of devices and coatings to significantly lower the drug detection limit and may also have an impact on our understanding of antibiotic drug action in bacteria.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    et al. Translating biomolecular recognition into nanomechanics. Science 288, 316–318 (2000).

  2. 2.

    et al. Multiple label-free biodetection and quantitative DNA-binding assays on a nanomechanical cantilever array. Proc. Natl Acad. Sci. USA 99, 9783–9788 (2002).

  3. 3.

    , , & Micromechanical detection of proteins using aptamer-based receptor molecules. Anal. Chem. 76, 3194–3198 (2004).

  4. 4.

    et al. Highly sensitive polymer-based cantilever-sensors for DNA detection. Ultramicrosc. 105, 215–222 (2005).

  5. 5.

    et al. DNA molecular motor driven micromechanical cantilever arrays. J. Am. Chem. Soc. 127, 17054–17060 (2005).

  6. 6.

    et al. Investigating the molecular mechanisms of in-plane mechanochemistry on cantilever arrays. J. Am. Chem. Soc. 129, 601–609 (2007).

  7. 7.

    et al. A label-free immunosensor array using single-chain antibody fragments. Proc. Natl Acad. Sci. USA 102, 14587–14592 (2005).

  8. 8.

    et al. Rapid and label-free nanomechanical detection of biomarker transcripts in human RNA. Nature Nanotech. 1, 214–220 (2006).

  9. 9.

    Bioassay of prostate-specific antigen (PSA) using microcantilevers. Nature Biotechnol. 19, 856–860 (2001).

  10. 10.

    Advances in membrane receptor screening and analysis. J. Mol. Recognit. 17, 286–315 (2004).

  11. 11.

    et al. Binding of a dimeric derivative of vancomycin to L-Lys-D-Ala-D-Lactate in solution and at a surface. Chem. Biol. 121, 353–359 (1999).

  12. 12.

    , , & Using surface plasmon resonance to study the binding of vancomycin and its dimer to self-assembled monolayers presenting D-Ala-D-Ala. J. Am. Chem. Soc. 121, 2629–2630 (1999).

  13. 13.

    , , & Binding of vancomycin group antibiotics to D-alanine and D-lactate presenting self-assembled monolayers. Bioorg. Med. Chem. 8, 2609–2616 (2000).

  14. 14.

    et al. Ultrahigh density, high-data-rate NEMS-based AFM data storage system. Microelec. Eng. 46, 11–14 (1999).

  15. 15.

    , , , & Wafer-scale microdevice transfer/interconnect: Its application in an AFM-based data-storage system. J. MEMS 13, 895–901 (2004).

  16. 16.

    The glycopeptide story—How to kill the deadly ‘superbugs’. Nat. Prod. Rep. 13, 469–477 (1996).

  17. 17.

    , , & An analysis of the origins of a cooperative binding energy of dimerization. Science 280, 711–714 (1998).

  18. 18.

    & The structure and mode of action of glycopeptides antibiotics of the vancomycin group. Annu. Rev. Microbiol. 38, 339–357 (1984).

  19. 19.

    , , & Glycopeptide and lipoglycopeptide antibiotics. Chem. Rev. 105, 425–448 (2005).

  20. 20.

    Specificity of combination between mucopeptide precursors and vancomycin or ristomycin. Biochem. J. 111, 195–205 (1969).

  21. 21.

    & Modifications of acyl-d-alanyl-d-alanine terminus affecting complex-formation with vancomycin. Biochem. J. 123, 773–787 (1971).

  22. 22.

    Changing patterns of infectious diseases. Nature 406, 762–767 (2000).

  23. 23.

    Molecular mechanisms that confer antibacterial drug resistance. Nature 406, 775–781 (2000).

  24. 24.

    The crisis in antibiotic resistance. Science 257, 1064–1073 (1992).

  25. 25.

    et al. Molecular-basis for vancomycin resistance in Enterococcus faecium BM4147—biosynthesis of a depsipeptide peptidoglycan precursor by vancomycin resistance proteins VanH and VanA. Biochemistry 30, 10408–10415 (1991).

  26. 26.

    A call to arms. Nature Rev. Drug Discov. 6, 8–12 (2007).

  27. 27.

    et al. Vancomycin derivatives that inhibit peptidoglycan biosynthesis without binding D-Ala-D-Ala. Science 284, 507–510 (1999).

  28. 28.

    & Self-assembled organic monolayers—model systems for studying adsorption of proteins at surfaces. Science 252, 1164–1167 (1991).

  29. 29.

    & Adsorption of proteins onto surfaces containing end-attached oligo(ethylene oxide)—a model system using self-assembled monolayers. J. Am. Chem. Soc. 115, 10714–10721 (1993).

  30. 30.

    , , & A strategy for the generation of surfaces presenting ligands for studies of binding based on an active ester as a common reactive intermediate: A surface plasmon resonance study. Anal. Chem. 71, 777–790 (1999).

  31. 31.

    The tension of metallic films deposited by electrolysis. Proc. R. Soc. Lond. A 82, 172–175 (1909).

  32. 32.

    Calculated elastic constants for stress problems associated with semiconductor devices. J. Appl. Phys. 44, 534–535 (1973).

  33. 33.

    et al. Pharmacokinetics of vancomycin: Observations in 28 patients and dosage recommendations. Antimicrob. Agents Chemother. 22, 391–394 (1982).

  34. 34.

    & Introduction to Percolation Theory 2nd edn (Taylor and Francis, 1991).

  35. 35.

    Experimental study of the elastic properties of a percolating system. Phys. Rev. Lett. 53, 2028–2030 (1984).

  36. 36.

    & Fracture behaviour of a solid with random porosity. Phys. Rev. Lett. 56, 2509–2512 (1986).

  37. 37.

    , , , & Physics of nanomechanical biosensing on cantilever arrays. Adv. Mater. (in the press).

  38. 38.

    , & Phase behaviour of two-component self- assembled monolayers of alkanethiolates on gold. J. Phys. Chem. 98, 563–571 (1994).

  39. 39.

    et al. Phase separation within a binary self-assembled monolayer on Au{111} driven by an amide-containing alkanethiol. J. Phys. Chem. B 105, 1119–1122 (2001).

Download references

Acknowledgements

We acknowledge funding from the Engineering and Physical Sciences Research Council (Speculative Engineering funding programme, J.W.N, R.McK., G.A., M.W., D.Z.), Interdisciplinary Research Centre in Nanotechnology (Cambridge, UCL and Bristol, M.W., J.W.N., R.McK.), the Royal Society (R.McK., J.W.N.), Biotechnology and Biological Science Research Council (R.McK.), the Wolfson Foundation (G.A.) and Cancer Research UK (A.D.B.). We thank E. Smith (University of Nottingham EPSRC XPS Open Access scheme) and J. and R. Galbraith (UCL Statistics) for helpful discussions.

Author information

Author notes

    • Joseph Wafula Ndieyira
    •  & Moyu Watari

    These authors contributed equally to this work.

Affiliations

  1. London Centre for Nanotechnology and Departments of Medicine and Physics, University College London, 17–19 Gordon Street, London WC1H 0AH, UK

    • Joseph Wafula Ndieyira
    • , Moyu Watari
    • , Alejandra Donoso Barrera
    • , Manuel Vögtli
    • , Torsten Strunz
    • , Mike A. Horton
    • , Gabriel Aeppli
    •  & Rachel A. McKendry
  2. Jomo Kenyatta University of Agriculture and Technology, Department of Chemistry, PO Box 62000, Nairobi, Kenya

    • Joseph Wafula Ndieyira
  3. Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK

    • Dejian Zhou
    • , Matthew Batchelor
    •  & Chris Abell
  4. School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK

    • Dejian Zhou
  5. Institute for Molecular Bioscience, The University of Queensland, Brisbane 4072, Australia

    • Matthew A. Cooper
  6. School of Chemistry, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK

    • Trevor Rayment

Authors

  1. Search for Joseph Wafula Ndieyira in:

  2. Search for Moyu Watari in:

  3. Search for Alejandra Donoso Barrera in:

  4. Search for Dejian Zhou in:

  5. Search for Manuel Vögtli in:

  6. Search for Matthew Batchelor in:

  7. Search for Matthew A. Cooper in:

  8. Search for Torsten Strunz in:

  9. Search for Mike A. Horton in:

  10. Search for Chris Abell in:

  11. Search for Trevor Rayment in:

  12. Search for Gabriel Aeppli in:

  13. Search for Rachel A. McKendry in:

Contributions

J.W.N. and M.W. contributed equally to this manuscript. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Rachel A. McKendry.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Information

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nnano.2008.275

Further reading