Integrated proteomic analysis of a bacterial pathogen -

Campylobacter jejuni

The bacterium Campylobacter jejuni is an important human pathogen with poorly understood mechanisms of virulence and colonization. To facilitate studies of this organism, the Finley lab and the Michigan Proteome Consortium set out to generate a proteome-wide protein interaction map for C. jejuni using the yeast two-hybrid system, to determine protein expression profiles by MS-based proteome mapping, and to generate clone resources for C. jejuni researchers.

C. jejuni as a model organism for proteome analysis.  C. jejuni, a microaerophilic Gram-negative bacterium, is a food-borne pathogen and a major cause of bacterial gastroenteritis throughout the world.  In the United States, the number of cases of Campylobacter infections (80-90% of which are caused by C. jejuni) significantly surpasses Escherichia coli, Shigella, and Salmonella species.  It has been estimated that each year one out of every 100 individuals in the United States and Great Britain develops Campylobacter infections.  C. jejuni infection might also be linked to Guillain-Barré Syndrome, a neurological disease that results in neuromuscular paralysis.

The genome sequence of C. jejuni NCT11168 has been determined.  The genome is 1.6 mb with 1,654 predicted proteins.  22.2% of the predicted proteins have no similarity to proteins with known functions, while the sequence of many of the others provides only limited clues about function. 

Results

A clone set for expressing C. jejuni proteins fused to an N-terminal GST-EK-6His tag in E. coli was constructed by recombination in E. coli. Clones are available from Source Bioscience. The clone set is described in:

High-throughput cloning of Campylobacter jejuni ORfs by in vivo recombination in Escherichia coli. Parrish JR, Limjindaporn T, Hines JA, Liu J, Liu G, Finley RL Jr.. J Proteome Res, 3(3):582-6, 2004. [Full text].

A large-scale yeast two-hybrid screen was conducted to detect binary protein-protein inetarctions for C. jejuni proteins. The data is freely available and can be visualized and downloaded from here. The data is described in:

Parrish JR, Yu J, Liu G, Hines JA, Chan JE, Mangiola BA, Zhang H, Pacifico S, Fotouhi F, Dirita VJ, Ideker T, Andrews P, Finley RL Jr. A proteome-wide protein interaction map for Campylobacter jejuni. Genome Biol. 2007 Jul 5;8(7):R130. PMID: 17615063.

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