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Applied and Environmental Microbiology, June 2003, p. 3368-3376, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3368-3376.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Development of a Listeria monocytogenes EGDe Partial Proteome Reference Map and Comparison with the Protein Profiles of Food Isolates

Manilduth Ramnath,^1,2 K. Björn Rechinger,^2, Lothar Jänsch,³ John W. Hastings,¹ Susanne Knøchel,² and Anne Gravesen^2*

Department of Biochemistry, University of Stellenbosch, 7602 Matieland, South Africa,¹ Department of Dairy and Food Science, Centre of Advanced Food Studies, LMC, The Royal Veterinary and Agricultural University, DK-1958 Frederiksberg C, Denmark,² Department of Cell Biology, German Research Center for Biotechnology, Braunschweig D-38124, Germany³

Received 13 September 2002/ Accepted 20 March 2003

A partially annotated proteome reference map of the food pathogen Listeria monocytogenes was developed for exponentially growing cells under standardized, optimal conditions by using the sequenced strain EGDe (serotype 1/2a) as a model organism. The map was developed by using a reproducible total protein extraction and two-dimensional (2-D) polyacrylamide gel electrophoresis analysis procedure, and it contained 33 identified proteins representing the four main protein functional classes. In order to facilitate analysis of membrane proteins, a protein compartmentalization procedure was assessed. The method used provided partial fractionation of membrane and cytosolic proteins. The total protein 2-D profiles of three serotype 1/2a strains and one serotype 1/2b strain isolated from food were compared to the L. monocytogenes EGDe proteome. An average of 13% of the major protein spots in the food strain proteomes were not matched in the strain EGDe proteome. The variation was greater for the less intense spots, and on average 28% of these spots were not matched. Two of the proteins identified in L. monocytogenes EGDe were missing in one or more of the food isolates. These two proteins were proteins involved in the main glycolytic pathway and in metabolism of coenzymes and prosthetic groups. The two corresponding genes were found by PCR amplification to be present in the four food isolates. Our results show that the L. monocytogenes EGDe reference map is a valuable starting point for analyses of strains having various origins and could be useful for analyzing the proteomes of different isolates of this pathogen.

Present address: Foss Electric A/S, DK-3400 Hillerød, Denmark.