Elsevier

Veterinary Microbiology

Volume 90, Issues 1–4, 20 December 2002, Pages 593-603
Veterinary Microbiology

Brucella proteomes—a review

https://doi.org/10.1016/S0378-1135(02)00239-0Get rights and content

Abstract

The proteomes of selected Brucella spp. have been extensively analyzed by utilizing current proteomic technology involving 2-DE and MALDI-MS. In Brucella melitensis, more than 500 proteins were identified. The rapid and large-scale identification of proteins in this organism was accomplished by using the annotated B. melitensis genome which is now available in the GenBank. Coupled with new and powerful tools for data analysis, differentially expressed proteins were identified and categorized into several classes. A global overview of protein expression patterns emerged, thereby facilitating the simultaneous analysis of different metabolic pathways in B. melitensis. Such a global characterization would not have been possible by using time consuming and traditional biochemical approaches. The era of post-genomic technology offers new and exciting opportunities to understand the complete biology of different Brucella species.

Introduction

Brucella melitensis is one of the organisms whose genome has been completely sequenced, closed, and annotated (DelVecchio et al., 2002). Although many aspects of its biology remain to be understood, the sequencing and annotation of its genome paved the way for a highly comprehensive and rapid analysis of its proteome. Proteome is defined as the entire set of proteins expressed at a specific time and under defined environmental conditions. This post-genomic approach of proteomics, relies heavily on two-dimensional gel electrophoresis (2-DE) for protein separation and on matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) for high throughput protein identification. Because the annotated B. melitensis genome has been available for some time, a rapid global analysis of its proteome using the Mascot search engine was initiated (Wagner et al., 2002). Computer-assisted comparison of hundreds of B. melitensis proteins from different strains resulted in categorizing levels of differentially expressed proteins into several classes (Eschenbrenner et al., 2002). It is anticipated that ultimately, the proteome data generated using these powerful techniques will provide answers to many interesting aspects of Brucella biology including virulence (see Ugalde, 1999 for a review), survival inside macrophages (O’Callaghan et al., 1999, Foulongne et al., 2000) and host preference (Corbel and Brinley-Morgan, 1984, Corbel, 1997, Enright, 1990). The intent of this review is to survey previous and current proteomics studies on Brucella spp. with emphasis on recent work on the global characterization of the B. melitensis proteome.

Section snippets

Early protein studies

Early studies have used one-dimensional SDS-PAGE and/or immunoblotting to describe differential sets of proteins expressed by various Brucella species (Morris, 1973, Santos et al., 1984, Gamazo et al., 1989, Brooks-Worrell and Splitter, 1992, Hill and Cook, 1994). In general, these 1-D PAGE studies revealed that protein band patterns were specific to each individual species. Similarly, differences between vaccine and virulent strains have been described (Verstreate et al., 1982, Tabatabai and

Global analysis of B. melitensis proteome

Using whole-cell protein extracts of laboratory-grown B. melitensis strain 16M, Wagner et al. (2002) examined the proteome using a series of 2-DE gel electrophoresis of overlapping narrow range IPG strips from pH 3.5 to 11.0. A total of 883 non-redundant protein spots were detected using SYPRO® Ruby protein stain. To date, 557 protein spots were identified by peptide mass fingerprinting which corresponded to 232 discrete ORFs, or 7.3% of the predicted 3197 ORFs of the 16M genome. A

Proteomes of vaccine strains

An attenuated strain of B. melitensis, called Rev 1, was developed in 1957 (Elberg and Faunce, 1957). It is considered a highly effective vaccine in the control of brucellosis in small ruminants in many countries (Blasco, 1997). A comprehensive proteome analysis of the laboratory-grown vaccine strain Rev 1 was reported by Eschenbrenner et al. (2002). Using the virulent strain 16M as reference, comparative analysis revealed that overall the 16M and Rev 1 proteomes are highly similar to each

B. melitensis versus B. abortus proteomes

The genomes of various Brucella species i.e. B. abortus, B. canis, B. melitensis, B. neotomae, B. ovis and B. suis are quite similar. DNA-DNA hybridization studies which suggested greater than 87% interspecies similarity (Verger et al., 1985) and pulsed field gel electrophoresis-separated PacI and SpeI macrorestriction maps (Michaux-Charachon et al., 1997) both support the classification of Brucella as a monospecific genus. However, the current classification scheme that is widely used

Conclusions and future directions

Genomic and proteomic technology have revolutionized the way we design and conduct current biological experiments. Our ability to analyze rapidly hundreds of expressed proteins and identify which gene encodes a specific protein generates vast amount of information essential in examining different but interrelated pathways within the organism. The data generated present a global but temporal view of metabolism. We have used the “proteome from the genome” approach in characterizing differentially

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