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

Juliane Düvel, Daniela Bertinetti, Stefan Möller, Frank Schwede, Michael Morr, Josef Wissing, Lena Radamm, Bastian Zimmermann, Hans-Gottfried Genieser, Lothar Jänsch, Friedrich W Herberg, Susanne Häussler

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.

OriginalsprogEngelsk
TidsskriftJournal of Microbiological Methods
Vol/bind88
Udgave nummer2
Sider (fra-til)229-36
Antal sider8
ISSN0167-7012
DOI
StatusUdgivet - feb. 2012
Udgivet eksterntJa

Fingeraftryk

Dyk ned i forskningsemnerne om 'A chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa'. Sammen danner de et unikt fingeraftryk.

Citationsformater