Quantitative determination of cyclic diguanosine monophosphate concentrations in nucleotide extracts of bacteria by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry.

2.50
Hdl Handle:
http://hdl.handle.net/10033/64253
Title:
Quantitative determination of cyclic diguanosine monophosphate concentrations in nucleotide extracts of bacteria by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry.
Authors:
Simm, Roger; Morr, Michael; Remminghorst, Uwe; Andersson, Mats; Römling, Ute
Abstract:
The 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.
Affiliation:
Department of Microbiology, Tumor and Cell Biology, Karolinska Institute, Stockholm, Sweden.
Citation:
Quantitative 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.
Journal:
Analytical biochemistry
Issue Date:
1-Mar-2009
URI:
http://hdl.handle.net/10033/64253
DOI:
10.1016/j.ab.2008.12.013
PubMed ID:
19135022
Type:
Article
Language:
en
ISSN:
1096-0309
Appears in Collections:
Publications of Dept. Cell Biology (ZB)

Full metadata record

DC FieldValue Language
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.en
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

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