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Improvement of existing anthrax vaccines, which are currently based on the administration of Protective Antigen (the highly immunogenic nontoxic subunit of the bacterial toxin) may entail other bacterial immunogenic elements, part of which are predicted to reside on the surface of bacterial cells. In the present study, membranal proteins extracted from a stationary\u2010phase culture of a nonvirulent <jats:italic>B. anthracis<\/jats:italic> strain, devoid of the native virulence plasmids pXO1 and pXO2, were separated by two\u2010dimensional electrophoresis (2\u2010DE) and a characteristic protein map was defined. The proteomic analysis allowed matix\u2010assisted laser desorption\/ionization\u2010time of flight mass spectrometry\u2010assisted identification of 86 protein spots which represent the product of 30 individual open reading frames (ORF). Among these, a prevalent class of proteins was the <jats:italic>S<\/jats:italic>\u2010layer proteins (which were found to represent more than 75% of the <jats:italic>B. anthracis<\/jats:italic> membranal fraction) and proteins containing <jats:italic>S<\/jats:italic>\u2010layer homology (SLH)\u2010membranal localization domains. Five novel SLH proteins, previously inferred only from bioinformatic ORF analysis (draft genome sequence), were identified and one was shown to be a highly abundant membranal protein. Western blots of the 2\u2010DE gels were probed with sera from convalescent rabbits and guinea pigs infected with virulent <jats:italic>B. anthracis<\/jats:italic> (Vollum strain). This analysis revealed that <jats:italic>B. anthracis<\/jats:italic> immune animals exhibit antibodies against at least 14 distinct membranal proteins present in the 2\u2010DE map, establishing that these proteins are expressed <jats:italic>in vivo<\/jats:italic> and are able to elicit an immune response. The identification of the protein components of the <jats:italic>B. anthracis<\/jats:italic> membranal fraction, as well as the establishment of their potential immunogenicity, underscore the strength of the proteomic approach for identifying molecules which may serve for further analysis of immune and protective abilities.<\/jats:p>","DOI":"10.1002\/pmic.200300575","type":"journal-article","created":{"date-parts":[[2004,2,26]],"date-time":"2004-02-26T15:35:05Z","timestamp":1077809705000},"page":"677-691","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":43,"title":["Identification of chromosomally encoded membranal polypeptides of <b><i>Bacillus anthracis<\/i><\/b> by a proteomic analysis: Prevalence of proteins containing <b><i>S<\/i><\/b>\u2010layer homology domains"],"prefix":"10.1002","volume":"4","author":[{"given":"Theodor","family":"Chitlaru","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Naomi","family":"Ariel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anat","family":"Zvi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Menahem","family":"Lion","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Baruch","family":"Velan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Avigdor","family":"Shafferman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eytan","family":"Elhanany","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2004,2,19]]},"reference":[{"key":"e_1_2_1_2_2","first-page":"543","volume-title":"Anthrax Toxins","author":"Leppla S. 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