A cyclic dinucleotide containing 2-aminopurine is a general fluorescent sensor for c-di-GMP and 3',3'-cGAMP.

Abstract

Cyclic dinucleotides have emerged as second messengers that regulate diverse processes in bacteria, as well as regulating the production of type I interferons in metazoans. Fluorescent sensors for these important second messengers are highly sought-after for high-throughput inhibitor discovery, yet most sensors reported to date are not amenable for high-throughput screening purposes. Herein, we demonstrate that a new analog, 3',3'-cG(d2AP)MP, which is a 2-aminopurine (2AP)-containing cyclic dinucleotide, self-associates in the presence of Mn(2+) with an association constant of 120,000 M(-1). 3'3'-cG(d2AP)MP can also form a heterodimer with cGAMP, activator of immune regulator, STING, or the bacterial biofilm regulator, c-di-GMP in the presence of Mn(II). Upon dimer formation, the fluorescence of 3',3'-cG(d2AP)MP is quenched and this provides a convenient method to monitor the enzymatic processing of both DGC and PDE enzymes, opening up several opportunities for the discovery of inhibitors of nucleotide signaling.

• Authors: Benjamin T Roembke, Jie Zhou, Yue Zheng, David Sayre, Allan Lizardo, Laurentee Bernard, Herman O Sintim
• Journal: Molecular bioSystems (Mol Biosyst) Vol. 10 Issue 6 Pg. 1568-75 (Jun 2014) ISSN: 1742-2051 [Electronic] England
• PMID: 24705858 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Molecular Probes
• Nucleotides, Cyclic
• cyclic guanosine monophosphate-adenosine monophosphate
• 2-Aminopurine
• Cyclic GMP

Topics

• 2-Aminopurine (metabolism)
• Biofilms
• Cyclic GMP (analogs & derivatives, chemistry)
• Fluorescence
• Molecular Probes (chemistry)
• Nucleotides, Cyclic (chemistry)
• Second Messenger Systems
• Signal Transduction