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c-di-GMP Signaling pp 431–453Cite as

Discovering Selective Diguanylate Cyclase Inhibitors: From PleD to Discrimination of the Active Site of Cyclic-di-GMP Phosphodiesterases

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1657))

Abstract

One of the most important signals involved in controlling biofilm formation is represented by the intracellular second messenger 3′,5′-cyclic diguanylic acid (c-di-GMP). Since the pathways involved in c-di-GMP biosynthesis and breakdown are found only in bacteria, targeting c-di-GMP metabolism represents an attractive strategy for the development of biofilm-disrupting drugs. Here, we present the workflow required to perform a structure-based design of inhibitors of diguanylate cyclases, the enzymes responsible for c-di-GMP biosynthesis. Downstream of the virtual screening process, detailed in the first part of the chapter, we report the step-by-step protocols required to test the positive hits in vitro and to validate their selectivity, thus minimizing possible off-target effects.

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Author information

Author notes

  1. S. Rinaldo and G. Giardina contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemical Sciences, Istituto Pasteur Italia-Fondazione Cenci Bolognetti, Sapienza University of Rome, P.le Aldo Moro 5, Rome, 00185, Italy

    S. Rinaldo, G. Giardina, F. Mantoni, Alessio Paone & Francesca Cutruzzolà

  2. Department of Biology and Biotechnology “Charles Darwin”, Sapienza University of Rome, Rome, Italy

    A. Paiardini

Authors
  1. S. Rinaldo
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  2. G. Giardina
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  3. F. Mantoni
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  4. A. Paiardini
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  5. Alessio Paone
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  6. Francesca Cutruzzolà
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Corresponding author

Correspondence to Francesca Cutruzzolà .

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Editors and Affiliations

  1. Department of Biological Sciences, Binghamton University Department of Biological Sciences, Binghamton, New York, USA

    Karin Sauer

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Rinaldo, S., Giardina, G., Mantoni, F., Paiardini, A., Paone, A., Cutruzzolà, F. (2017). Discovering Selective Diguanylate Cyclase Inhibitors: From PleD to Discrimination of the Active Site of Cyclic-di-GMP Phosphodiesterases. In: Sauer, K. (eds) c-di-GMP Signaling. Methods in Molecular Biology, vol 1657. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7240-1_32

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  • DOI: https://doi.org/10.1007/978-1-4939-7240-1_32

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7239-5

  • Online ISBN: 978-1-4939-7240-1

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