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

Jie Zhou; Clement Opoku-Temeng; Herman O Sintim

doi:10.1007/978-1-4939-7240-1_19

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

Methods Mol Biol. 2017:1657:245-261. doi: 10.1007/978-1-4939-7240-1_19.

Authors

Jie Zhou^{1

2}, Clement Opoku-Temeng^{1

3

4}, Herman O Sintim^{5

6

7}

Affiliations

¹ Purdue Institute for Drug Discovery, Purdue University, 500 Oval Drive, West Lafayette, IN, 47907, USA.
² Department of Chemistry, Purdue University, West Lafayette, IN, 47907, USA.
³ Biochemistry Graduate Program, University of Maryland, College Park, MD, 20742, USA.
⁴ Department of Chemistry, Center for Drug Discovery, Purdue University, West Lafayette, IN, 47907, USA.
⁵ Purdue Institute for Drug Discovery, Purdue University, 500 Oval Drive, West Lafayette, IN, 47907, USA. hsintim@purdue.edu.
⁶ Department of Chemistry, Purdue University, West Lafayette, IN, 47907, USA. hsintim@purdue.edu.
⁷ Purdue Institute of Inflammation, Immunology and Infectious Disease, West Lafayette, IN, 47907, USA. hsintim@purdue.edu.

PMID: 28889299
DOI: 10.1007/978-1-4939-7240-1_19

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.

Keywords: 2-Aminopurine (2AP); Fluorescent bases; High-throughput screening; Inhibitor; Oligoribonuclease (Orn); c-di-GMP; c-di-GMP PDEs; c-di-GMP synthases.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, Non-U.S. Gov't

MeSH terms

2-Aminopurine* / chemistry
Cyclic GMP / analogs & derivatives*
Cyclic GMP / metabolism
Cyclic Nucleotide Phosphodiesterases, Type 6 / metabolism*
Enzyme Activation
Enzyme Assays / methods
Exoribonucleases
Fluorescent Dyes* / chemistry
Phosphoric Diester Hydrolases / metabolism*

Substances

Fluorescent Dyes
2-Aminopurine
bis(3',5')-cyclic diguanylic acid
Exoribonucleases
oligoribonuclease
Phosphoric Diester Hydrolases
Cyclic Nucleotide Phosphodiesterases, Type 6
Cyclic GMP