A chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa.

2.50
Hdl Handle:
http://hdl.handle.net/10033/244691
Title:
A chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa.
Authors:
Düvel, Juliane; Bertinetti, Daniela; Möller, Stefan; Schwede, Frank; Morr, Michael; Wissing, Josef; Radamm, Lena; Zimmermann, Bastian; Genieser, Hans-Gottfried; Jänsch, Lothar; Herberg, Friedrich W; Häussler, Susanne
Abstract:
In many bacteria, high levels of the ubiquitous second messenger c-di-GMP have been demonstrated to suppress motility and to promote the establishment of surface-adherent biofilm communities. While molecular mechanisms underlying the synthesis and degradation of c-di-GMP have been comprehensively characterized, little is known about how c-di-GMP mediates its regulatory effects. In this study, we have established a chemical proteomics approach to identify c-di-GMP interacting proteins in the opportunistic pathogen Pseudomonas aeruginosa. A functionalized c-di-GMP analog, 2'-aminohexylcarbamoyl-c-di-GMP (2'-AHC-c-di-GMP), was chemically synthesized and following its immobilization used to perform affinity pull down experiments. Enriched proteins were subsequently identified by high-resolution mass spectrometry. 2'-AHC-c-di-GMP was also employed in surface plasmon resonance studies to evaluate and quantify the interaction of c-di-GMP with its potential target molecules in vitro. The biochemical tools presented here may serve the identification of novel classes of c-di-GMP effectors and thus contribute to a better characterization and understanding of the complex c-di-GMP signaling network.
Affiliation:
Department of Cell Biology, Helmholtz Center for Infection Research, Inhoffenstr. 7, D-38124 Braunschweig, Germany.
Citation:
A chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa. 2012, 88 (2):229-36 J. Microbiol. Methods
Journal:
Journal of microbiological methods
Issue Date:
Feb-2012
URI:
http://hdl.handle.net/10033/244691
DOI:
10.1016/j.mimet.2011.11.015
PubMed ID:
22178430
Type:
Article
Language:
en
ISSN:
1872-8359
Appears in Collections:
publications of the division molecular bacteriology (MOBA)

Full metadata record

DC FieldValue Language
dc.contributor.authorDüvel, Julianeen_GB
dc.contributor.authorBertinetti, Danielaen_GB
dc.contributor.authorMöller, Stefanen_GB
dc.contributor.authorSchwede, Franken_GB
dc.contributor.authorMorr, Michaelen_GB
dc.contributor.authorWissing, Josefen_GB
dc.contributor.authorRadamm, Lenaen_GB
dc.contributor.authorZimmermann, Bastianen_GB
dc.contributor.authorGenieser, Hans-Gottfrieden_GB
dc.contributor.authorJänsch, Lotharen_GB
dc.contributor.authorHerberg, Friedrich Wen_GB
dc.contributor.authorHäussler, Susanneen_GB
dc.date.accessioned2012-09-18T14:16:26Z-
dc.date.available2012-09-18T14:16:26Z-
dc.date.issued2012-02-
dc.identifier.citationA chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa. 2012, 88 (2):229-36 J. Microbiol. Methodsen_GB
dc.identifier.issn1872-8359-
dc.identifier.pmid22178430-
dc.identifier.doi10.1016/j.mimet.2011.11.015-
dc.identifier.urihttp://hdl.handle.net/10033/244691-
dc.description.abstractIn many bacteria, high levels of the ubiquitous second messenger c-di-GMP have been demonstrated to suppress motility and to promote the establishment of surface-adherent biofilm communities. While molecular mechanisms underlying the synthesis and degradation of c-di-GMP have been comprehensively characterized, little is known about how c-di-GMP mediates its regulatory effects. In this study, we have established a chemical proteomics approach to identify c-di-GMP interacting proteins in the opportunistic pathogen Pseudomonas aeruginosa. A functionalized c-di-GMP analog, 2'-aminohexylcarbamoyl-c-di-GMP (2'-AHC-c-di-GMP), was chemically synthesized and following its immobilization used to perform affinity pull down experiments. Enriched proteins were subsequently identified by high-resolution mass spectrometry. 2'-AHC-c-di-GMP was also employed in surface plasmon resonance studies to evaluate and quantify the interaction of c-di-GMP with its potential target molecules in vitro. The biochemical tools presented here may serve the identification of novel classes of c-di-GMP effectors and thus contribute to a better characterization and understanding of the complex c-di-GMP signaling network.en_GB
dc.language.isoenen
dc.rightsArchived with thanks to Journal of microbiological methodsen_GB
dc.subject.meshBacterial Proteinsen_GB
dc.subject.meshCarrier Proteinsen_GB
dc.subject.meshCyclic GMPen_GB
dc.subject.meshProteomicsen_GB
dc.subject.meshPseudomonas aeruginosaen_GB
dc.subject.meshSignal Transductionen_GB
dc.subject.meshSurface Plasmon Resonanceen_GB
dc.titleA chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa.en
dc.typeArticleen
dc.contributor.departmentDepartment of Cell Biology, Helmholtz Center for Infection Research, Inhoffenstr. 7, D-38124 Braunschweig, Germany.en_GB
dc.identifier.journalJournal of microbiological methodsen_GB

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