Identification and Quantification of Cyclic Di-Guanosine Monophosphate and Its Linear Metabolites by Reversed-Phase LC-MS/MS

Heike Bähre; Volkhard Kaever

doi:10.1007/978-1-4939-7240-1_5

Identification and Quantification of Cyclic Di-Guanosine Monophosphate and Its Linear Metabolites by Reversed-Phase LC-MS/MS

Methods Mol Biol. 2017:1657:45-58. doi: 10.1007/978-1-4939-7240-1_5.

Authors

Heike Bähre¹, Volkhard Kaever²

Affiliations

¹ Research Core Unit Metabolomics, Institute of Pharmacology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany.
² Research Core Unit Metabolomics, Institute of Pharmacology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany. kaever.volkhard@mh-hannover.de.

PMID: 28889285
DOI: 10.1007/978-1-4939-7240-1_5

Abstract

Cyclic dinucleotides such as bis-(3',5')-cyclic dimeric guanosine monophosphate (3',3'-c-di-GMP) represent an important class of second messengers in bacteria and are involved in numerous (patho)physiological settings. Here, we describe a sensitive and specific quantification method for 3',3'-c-di-GMP by HPLC-coupled tandem mass spectrometry (LC-MS/MS). Additionally, linear 3',3'-c-di-GMP metabolites, i.e., 5'-phosphoguanylyl-3',5'-guanosine (pGpG) and 5'-guanosine monophosphate (5'-GMP), as well as cyclic guanosine monophosphate (3',5'-cGMP) and 3',3' c-di-GMP analogues (2',3'-c-di-GMP and 2',2'-c-di-GMP) can be simultaneously determined by this method.

Keywords: Cyclic di-GMP; HPLC; Tandem mass spectrometry.

MeSH terms

Bacteria / metabolism
Chromatography, Reverse-Phase* / methods
Chromatography, Reverse-Phase* / standards
Cyclic GMP / analogs & derivatives*
Cyclic GMP / chemistry
Cyclic GMP / metabolism
Metabolomics / methods
Second Messenger Systems
Tandem Mass Spectrometry* / methods
Tandem Mass Spectrometry* / standards

Substances

bis(3',5')-cyclic diguanylic acid
Cyclic GMP