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Fluorescent 2-Aminopurine c-di-GMP and GpG Analogs as PDE Probes

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Book cover c-di-GMP Signaling

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

Abstract

c-di-GMP is widely recognized as an important ubiquitous signaling molecule in bacteria. c-di-GMP phosphodiesterases (PDEs) regulate the intracellular concentration of c-di-GMP and some could be potential drug targets. Here, we describe a class of dinucleotide probes suitable for monitoring the enzymatic activities of c-di-GMP PDEs in real time. Such probes contain fluorescent nucleobases and can be readily cleaved by PDEs, resulting in a change in fluorescence. Fluorescent cyclic and linear dinucleotide probes could be used in diverse applications, such as confirming the activity of an expressed PDE or oligoribonuclease (Orns) or identifying inhibitors of PDEs or Orns using high-throughput screening formats.

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Acknowledgment

This work was funded by the National Science Foundation (CHE1307218 and CHE1636752), Purdue University. Plasmids were provided by Dr. Zhaoxun Liang (RocR plasmid), Dr. Ehud Banin (P. aeruginosa Orn plasmid), and Dr. Nicholas Dixon (E. coli and M. smegmatis Orn plasmids).

Author information

Authors and Affiliations

  1. Purdue Institute for Drug Discovery, Purdue University, 500 Oval Drive, West Lafayette, IN, 47907, USA

    Jie Zhou, Clement Opoku-Temeng & Herman O. Sintim

  2. Department of Chemistry, Purdue University, West Lafayette, IN, 47907, USA

    Jie Zhou & Herman O. Sintim

  3. Biochemistry Graduate Program, University of Maryland, College Park, MD, 20742, USA

    Clement Opoku-Temeng

  4. Purdue Institute of Inflammation, Immunology and Infectious Disease, West Lafayette, IN, 47907, USA

    Herman O. Sintim

  5. Department of Chemistry, Center for Drug Discovery, Purdue University, West Lafayette, IN, 47907, USA

    Clement Opoku-Temeng

Authors
  1. Jie Zhou
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  2. Clement Opoku-Temeng
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  3. Herman O. Sintim
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Corresponding author

Correspondence to Herman O. Sintim .

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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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Zhou, J., Opoku-Temeng, C., Sintim, H.O. (2017). Fluorescent 2-Aminopurine c-di-GMP and GpG Analogs as PDE Probes. 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_19

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  • DOI: https://doi.org/10.1007/978-1-4939-7240-1_19

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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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