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.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Düvel J,Bertinetti D,Möller S,Schwede F,Morr M,Wissing J,Radamm L,Zimmermann B,Genieser HG,Jänsch L,Herberg FW,Häussler Sdoi
10.1016/j.mimet.2011.11.015subject
Has Abstractpub_date
2012-02-01 00:00:00pages
229-36issue
2eissn
0167-7012issn
1872-8359pii
S0167-7012(11)00407-6journal_volume
88pub_type
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