کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2090323 1081494 2012 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
پیش نمایش صفحه اول مقاله
A chemical proteomics approach to identify c-di-GMP binding proteins in Pseudomonas aeruginosa
چکیده انگلیسی

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.


► We immobilize functionalized c-di-GMP on a solid support.
► This affinity matrix serves the isolation of c-di-GMP binding proteins.
► MS-identified proteins harbor known but also new c-di-GMP binding motifs.
► We establish surface plasmon resonance assays to validate c-di-GMP binding.
► The biochemical tools will help to understand the complex c-di-GMP signaling system.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Microbiological Methods - Volume 88, Issue 2, February 2012, Pages 229–236
نویسندگان
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