Quantitation of cyclic dinucleotides by reversed-phase LC-MS/MS
© Spangler et al; licensee BioMed Central Ltd. 2009
Published: 11 August 2009
Cyclic nucleotides function as second messengers in all kingdoms of life. Recently, cyclic dinucleotides (c-di-NMPs) have been identified as bacterial signaling molecules. Cyclic diguanosine monophosphate (c-di-GMP) plays a major role in the modulation of bacterial surface components. Synthesis and degradation of c-di-GMP is controlled by so-called GGDEF (diguanylate cyclase activity) and EAL (phosphodiesterase activity) domains, respectively. These domains influence the sessility and motility of bacteria via regulation of biosynthesis of exopolysaccharides (e.g. cellulose) and fimbriae. Thus, the c-di-GMP level affects the formation of biofilms that represent the predominant form in which bacteria exist in the natural environment. The progress and persistence of many infectious diseases is based on biofilm formation by bacterial pathogens.
We will apply the established method for the quantitation of c-di-NMPs in various Pseudomonas aeruginosa strains and aim at determining the enzymatic activity of diguanylate cyclase.
The authors thank Dr. H.-G. Genieser and Dr. F. Schwede (BioLog, Bremen, Germany) for providing cyclic dinucleotide standards.
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