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dc.contributor.authorSimm, Roger
dc.contributor.authorMorr, Michael
dc.contributor.authorRemminghorst, Uwe
dc.contributor.authorAndersson, Mats
dc.contributor.authorRömling, Ute
dc.date.accessioned2009-04-03T12:53:23Z
dc.date.available2009-04-03T12:53:23Z
dc.date.issued2009-03-01
dc.identifier.citationQuantitative determination of cyclic diguanosine monophosphate concentrations in nucleotide extracts of bacteria by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. 2009, 386 (1):53-8 Anal. Biochem.en
dc.identifier.issn1096-0309
dc.identifier.pmid19135022
dc.identifier.doi10.1016/j.ab.2008.12.013
dc.identifier.urihttp://hdl.handle.net/10033/64253
dc.description.abstractThe physiological response to small molecules (secondary messengers) is the outcome of a delicate equilibrium between biosynthesis and degradation of the signal. Cyclic diguanosine monophosphate (c-di-GMP) is a novel secondary messenger present in many bacteria. It has a complex cellular metabolism whereby usually more than one enzyme synthesizing and degrading c-di-GMP is encoded by a bacterial genome. To assess the in vivo conditions of c-di-GMP signaling, we developed a high-performance liquid chromatography (HPLC)-mass spectrometry-based method to detect c-di-GMP with high sensitivity and to quantify the c-di-GMP concentration in the bacterial cell as described here in detail. We successfully used the methodology to determine and compare the c-di-GMP concentrations in bacterial species such as Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa, and Vibrio cholerae. We describe the use of the methodology to assess the change in c-di-GMP concentration during the growth phase and the contribution of a point mutation in S. typhimurium to the overall cellular c-di-GMP concentration.
dc.language.isoenen
dc.subject.meshBacteriaen
dc.subject.meshCyclic GMPen
dc.subject.meshMethodsen
dc.subject.meshNucleotidesen
dc.subject.meshResearch Designen
dc.subject.meshSecond Messenger Systemsen
dc.subject.meshTandem Mass Spectrometryen
dc.titleQuantitative determination of cyclic diguanosine monophosphate concentrations in nucleotide extracts of bacteria by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry.en
dc.typeArticleen
dc.contributor.departmentDepartment of Microbiology, Tumor and Cell Biology, Karolinska Institute, Stockholm, Sweden.en
dc.identifier.journalAnalytical biochemistryen
refterms.dateFOA2018-06-12T18:05:35Z
html.description.abstractThe physiological response to small molecules (secondary messengers) is the outcome of a delicate equilibrium between biosynthesis and degradation of the signal. Cyclic diguanosine monophosphate (c-di-GMP) is a novel secondary messenger present in many bacteria. It has a complex cellular metabolism whereby usually more than one enzyme synthesizing and degrading c-di-GMP is encoded by a bacterial genome. To assess the in vivo conditions of c-di-GMP signaling, we developed a high-performance liquid chromatography (HPLC)-mass spectrometry-based method to detect c-di-GMP with high sensitivity and to quantify the c-di-GMP concentration in the bacterial cell as described here in detail. We successfully used the methodology to determine and compare the c-di-GMP concentrations in bacterial species such as Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa, and Vibrio cholerae. We describe the use of the methodology to assess the change in c-di-GMP concentration during the growth phase and the contribution of a point mutation in S. typhimurium to the overall cellular c-di-GMP concentration.


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