%0 Journal Article %1 duvel2011chemical %A Duvel, J. %A Bertinetti, D. %A Muller, S. %A Schwede, F. %A Morr, M. %A Wissing, J. %A Radamm, L. %A Zimmermann, B. %A Genieser, H.-G. %A Jansch, L. %A Herberg, F. W. %A Haussler, S. %D 2012 %J Journal of Microbiological Methods %K herberg myown %N 2 %P 229-36 %R 10.1016/j.mimet.2011.11.015 %T A chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa %U http://www.ncbi.nlm.nih.gov/pubmed/22178430 %V 88 %X 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.

@article{duvel2011chemical, 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.}, added-at = {2012-01-09T15:15:35.000+0100}, author = {Duvel, J. and Bertinetti, D. and Muller, S. and Schwede, F. and Morr, M. and Wissing, J. and Radamm, L. and Zimmermann, B. and Genieser, H.-G. and Jansch, L. and Herberg, F. W. and Haussler, S.}, biburl = {https://www.bibsonomy.org/bibtex/2aed75fb80ca3bad8b42453c73c0af3f2/biochemie}, description = {A chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa 10.1016/j.mimet.2011.11.015 : Journal of Microbiological Methods | ScienceDirect.com}, doi = {10.1016/j.mimet.2011.11.015}, interhash = {bb120e3368110154fe2ab9cd630ba0f8}, intrahash = {aed75fb80ca3bad8b42453c73c0af3f2}, issn = {0167-7012}, journal = {Journal of Microbiological Methods}, keywords = {herberg myown}, number = 2, pages = {229-36 }, timestamp = {2012-07-24T09:38:40.000+0200}, title = {A chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa}, url = {http://www.ncbi.nlm.nih.gov/pubmed/22178430}, volume = 88, year = 2012 }