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Specific secretion machineries which are responsible for the subfraction of secreted proteins (soluble supernatant proteins [SSPs]) and the production of bacterial outer membrane vesicles (OMVs) both contribute to the protein composition of the extracellular milieu of this lung pathogen. Here we present comprehensive proteome reference maps for both SSPs and OMVs. Protein identification and assignment analyses revealed a total of 181 supernatant proteins, 107 of which were specific to the SSP fraction and 33 of which were specific to OMVs. A functional classification showed that a large proportion of the identified OMV proteins are involved in the pathogenesis of Legionnaires' disease. Zymography and enzyme assays demonstrated that the SSP and OMV fractions possess proteolytic and lipolytic enzyme activities which may contribute to the destruction of the alveolar lining during infection. Furthermore, it was shown that OMVs do not kill host cells but specifically modulate their cytokine response. Binding of immunofluorescently stained OMVs to alveolar epithelial cells, as visualized by confocal laser scanning microscopy, suggested that there is delivery of a large and complex group of proteins and lipids in the infected tissue in association with OMVs. On the basis of these new findings, we discuss the relevance of protein sorting and compartmentalization of virulence factors, as well as environmental aspects of the vesicle-mediated secretion.\n                  <\/jats:p>","DOI":"10.1128\/iai.01396-07","type":"journal-article","created":{"date-parts":[[2008,2,4]],"date-time":"2008-02-04T20:46:01Z","timestamp":1202157961000},"page":"1825-1836","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":173,"title":["Proteomic Characterization of the Whole Secretome of\n                    <i>Legionella pneumophila<\/i>\n                    and Functional Analysis of Outer Membrane Vesicles"],"prefix":"10.1128","volume":"76","author":[{"given":"Frank","family":"Galka","sequence":"first","affiliation":[{"name":"Institut fu\u0308r Molekulare Infektionsbiologie, Julius-Maximilians-Universita\u0308t Wu\u0308rzburg, Ro\u0308ntgenring 11, D-97070 Wu\u0308rzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sun Nyunt","family":"Wai","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Umea\u030a University, S-901 87 Umea\u030a, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harald","family":"Kusch","sequence":"additional","affiliation":[{"name":"Institut fu\u0308r Mikrobiologie, Ernst-Moritz-Arndt-Universita\u0308t, F.-L.-Jahn-Str. 15, D-17487 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susanne","family":"Engelmann","sequence":"additional","affiliation":[{"name":"Institut fu\u0308r Mikrobiologie, Ernst-Moritz-Arndt-Universita\u0308t, F.-L.-Jahn-Str. 15, D-17487 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Hecker","sequence":"additional","affiliation":[{"name":"Institut fu\u0308r Mikrobiologie, Ernst-Moritz-Arndt-Universita\u0308t, F.-L.-Jahn-Str. 15, D-17487 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bernd","family":"Schmeck","sequence":"additional","affiliation":[{"name":"Medizinische Klinik m. 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