Measuring cyclic diguanylate (c-di-GMP)-specific phosphodiesterase activity using the MANT-c-di-GMP assay

Dorit Eli, Trevor E. Randall, Henrik Almblad, Joe J. Harrison, Ehud Banin

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations


The second messenger, cyclic diguanylate (c-di-GMP), regulates a variety of bacterial cellular and social behaviors. A key determinant of c-di-GMP levels in cells is its degradation by c-di-GMP-specific phosphodiesterases (PDEs). Here, we describe an assay to determine c-di-GMP degradation rates in vitro using 2′-O-(N′-methylanthraniloyl)-cyclic diguanylate (MANT-c-di-GMP). Additionally, a protocol for the production and purification of recombinant Pseudomonas aeruginosa RocR, a c-di-GMP-specific PDE that may serve as a control in MANT-c-di-GMP assays, is provided. The use of the fluorescent MANT-c-di-GMP analogue can deliver fundamental information about PDE function, and is suitable for identifying and investigating c-di-GMP-specific PDE activators and inhibitors.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages16
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media LLC 2017.


  • Cyclic diguanylate
  • MANT-c-di-GMP
  • Phosphodiesterase
  • RocR
  • pGpG


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