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Quantification of cyclic dinucleotides by reversed-phase LC-MS/MS
Heike Burhenne1, Volkhard Kaever
1Research Core Unit for Mass Spectrometry-Metabolomics, Institute of Pharmacology, Hannover Medical School, Hannover, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|May 18, 2013
Summary
This study introduces a sensitive HPLC-tandem mass spectrometry method for quantifying bacterial cyclic dinucleotides like c-di-AMP and c-di-GMP. This technique aids in understanding their role in plant-microbe interactions.
Area of Science:
- Microbiology
- Plant Science
- Analytical Chemistry
Background:
- Cyclic dinucleotides (CDNs) like bis-(3',5')-cyclic dimeric adenosine monophosphate (c-di-AMP) and bis-(3',5')-cyclic dimeric guanosine monophosphate (c-di-GMP) are crucial bacterial second messengers.
- Their synthesis and function in plants remain largely unexplored, despite potential roles in plant-microbe interactions such as phytopathogen and rhizobium symbiosis.
Purpose of the Study:
- To develop and validate a sensitive and specific quantification method for c-di-AMP and c-di-GMP.
- To enable simultaneous determination of related nucleotide metabolites.
Main Methods:
- High-Performance Liquid Chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS).
- Development of a method for sensitive and specific CDN detection.
Main Results:
- A highly sensitive and specific HPLC-MS/MS method for quantifying c-di-AMP and c-di-GMP was established.
- The method allows for the simultaneous detection of linear dinucleotide metabolites, mononucleotides, and cyclic mononucleotides.
Conclusions:
- The developed HPLC-MS/MS method provides a robust tool for analyzing CDNs and related nucleotides.
- This quantification method will facilitate research into the roles of CDNs in bacterial physiology and plant-associated microbial communities.
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