{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T08:44:33Z","timestamp":1792572273638,"version":"4.5.0"},"reference-count":49,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2005,1,25]],"date-time":"2005-01-25T00:00:00Z","timestamp":1106611200000},"content-version":"vor","delay-in-days":24,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteomics"],"published-print":{"date-parts":[[2005,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The severe pulmonary form of leptospirosis (SPFL) is an especially serious and rapid disease process characterized by alveolar hemorrhage and acute respiratory failure. The outer membrane of <jats:italic>Leptospira<\/jats:italic> facilitates direct interactions with the environs and likely contains important constituents involved during infection, transmission, survival, and adaptation to environmental conditions, including putative vaccinogen and diagnostic candidates. Outer membrane vesicles (OMVs) were purified by incubation in low\u2010pH citrate buffer, treatment in a French press, and centrifugation over a continuous sucrose gradient. OMVs characterized by two\u2010dimensional gel electrophoresis (2\u2010DE) contained the previously described outer membrane proteins OmpL1, Qlp42, LipL32, LipL41, LipL36 and Loa22. In addition, unknown, hypothetical and putative outer membrane proteins were identified. High\u2010performance liquid chromatography (HPLC) coupled with mass spectrometry and fraction collection (LC\u2010MS+) measured the intact mass profile of the major outer membrane protein, LipL32, and the putative lipoprotein Qlp42. In contrast to a predicted molecularmass of 27\u00a0653.5\u00a0Da for LipL32 after cleavage of its signal peptide, intact mass proteomics measured the mass as ranging from 28\u00a0468 to 28\u00a0583\u00a0Da, consistent with lipidation of LipL32. In contrast to a predicted molecular mass of 39.8\u00a0kDa for Qlp42, the actual mass was measured as 24\u00a0811 and 26\u00a0461\u00a0Da consistent with a 30\u00a0kDa doublet observed on sodium dodecyl sulfate\u2010polyacrylamide gel electrophoresis (SDS\u2010PAGE) gels and processing of the <jats:italic>N<\/jats:italic>\u2010terminus of the mature protein. These studies indicate that purified OMVs are highly compatible with proteomics technologies including 2\u2010DE and intact mass proteomics using LC\u2010MS+ that facilitates definition of actual molecular masses of intact outer membrane proteins, and heterogeneity associated with them.\u2201<\/jats:p>","DOI":"10.1002\/pmic.200400880","type":"journal-article","created":{"date-parts":[[2005,1,25]],"date-time":"2005-01-25T08:53:59Z","timestamp":1106643239000},"page":"144-152","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":74,"title":["Purification and proteomic analysis of outer membrane vesicles from a clinical isolate of <b><i>Leptospira interrogans<\/i><\/b> serovar Copenhageni"],"prefix":"10.1002","volume":"5","author":[{"given":"Jarlath E.","family":"Nally","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julian P.","family":"Whitelegge","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodrigo","family":"Aguilera","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martha M.","family":"Pereira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David R.","family":"Blanco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael A.","family":"Lovett","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2005,1,25]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.14.2.296-326.2001"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1097\/00001432-200110000-00005"},{"key":"e_1_2_1_4_2","volume-title":"Leptospira and Leptospirosis","author":"Faine S.","year":"1999"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1590\/S0102-311X2001000700014"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1086\/340615"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1086\/339942"},{"key":"e_1_2_1_8_2","doi-asserted-by":"publisher","DOI":"10.1007\/s100960000433"},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1067\/mem.2001.116399"},{"key":"e_1_2_1_10_2","doi-asserted-by":"publisher","DOI":"10.1093\/ije\/19.3.743"},{"key":"e_1_2_1_11_2","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1093\/milmed\/162.2.101","volume":"162","author":"Katz A. 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