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Surface-exposed outer membrane proteins (SE-OMPs) are crucial for bacterial\u2013host interactions. SE-OMPs locate and expose their epitope on cell surface where is easily accessed by host molecules. This study aimed to screen for surface-exposed proteins and their abundance profile of pathogenic <jats:italic>Leptospira interrogans<\/jats:italic> serovar Pomona. Two complementary approaches, surface biotinylation and surface proteolytic shaving, followed by liquid chromatography tandem-mass spectrometry <jats:italic>(<\/jats:italic>LC-MS\/MS) were employed to identify SE-OMPs of intact leptospires. For quantitative comparison, in-depth label-free analysis of SE-OMPs obtained from each method was performed using MaxQuant. The total number of proteins identified was 1,001 and 238 for surface biotinylation and proteinase K shaving, respectively. Among these, 39 were previously known SE-OMPs and 68 were predicted to be localized on the leptospiral surface. Based on MaxQuant analysis for relative quantification, six known SE-OMPs including EF- Tu, LipL21, LipL41, LipL46, Loa22, and OmpL36, and one predicted SE-OMPs, LipL71 were found in the 20 most abundant proteins, in which LipL41 was the highest abundant SE-OMP. Moreover, uncharacterized LIC14011 protein (LIP3228 ortholog in serovar Pomona) was identified as a novel predicted surface \u03b2b-OMP. High-abundance leptospiral SE-OMPs identified in this study may play roles in virulence and infection and are potential targets for development of vaccine or diagnostic tests for leptospirosis.<\/jats:p>","DOI":"10.1371\/journal.pntd.0009983","type":"journal-article","created":{"date-parts":[[2021,11,29]],"date-time":"2021-11-29T18:59:04Z","timestamp":1638212344000},"page":"e0009983","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":10,"title":["Surface proteomics and label-free quantification of Leptospira interrogans serovar Pomona"],"prefix":"10.1371","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8188-9934","authenticated-orcid":true,"given":"Teerasit","family":"Techawiwattanaboon","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Praparat","family":"Thaibankluay","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chahya","family":"Kreangkaiwal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Suwitra","family":"Sathean-Anan-Kun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6199-7420","authenticated-orcid":true,"given":"Prasong","family":"Khaenam","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiradej","family":"Makjaroen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Trairak","family":"Pisitkun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1781-7012","authenticated-orcid":true,"given":"Kanitha","family":"Patarakul","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2021,11,29]]},"reference":[{"issue":"12","key":"pntd.0009983.ref001","doi-asserted-by":"crossref","first-page":"757","DOI":"10.1016\/S1473-3099(03)00830-2","article-title":"Leptospirosis: a zoonotic disease of global importance","volume":"3","author":"AR Bharti","year":"2003","journal-title":"Lancet Infect Dis"},{"issue":"3\u20134","key":"pntd.0009983.ref002","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.vetmic.2009.03.012","article-title":"Leptospira and leptospirosis","volume":"140","author":"B Adler","year":"2010","journal-title":"Vet Microbiol"},{"issue":"9","key":"pntd.0009983.ref003","doi-asserted-by":"crossref","first-page":"e0003898","DOI":"10.1371\/journal.pntd.0003898","article-title":"Global Morbidity and Mortality of Leptospirosis: A Systematic Review","volume":"9","author":"F Costa","year":"2015","journal-title":"PLoS Negl Trop Dis"},{"issue":"12","key":"pntd.0009983.ref004","doi-asserted-by":"crossref","first-page":"2141","DOI":"10.3201\/eid2112.140659","article-title":"High Prevalence of Intermediate Leptospira spp. DNA in Febrile Humans from Urban and Rural Ecuador","volume":"21","author":"J Chiriboga","year":"2015","journal-title":"Emerg Infect Dis"},{"issue":"22","key":"pntd.0009983.ref005","doi-asserted-by":"crossref","first-page":"6074","DOI":"10.1128\/JB.01119-12","article-title":"Leptospiral outer membrane protein microarray, a novel approach to identification of host ligand-binding proteins","volume":"194","author":"M Pinne","year":"2012","journal-title":"J Bacteriol"},{"issue":"2","key":"pntd.0009983.ref006","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1111\/1574-6968.12349","article-title":"Leptospiral extracellular matrix adhesins as mediators of pathogen-host interactions","volume":"352","author":"ML Vieira","year":"2014","journal-title":"FEMS Microbiol Lett"},{"key":"pntd.0009983.ref007","first-page":"2","article-title":"Bacterial surface proteins and vaccines","author":"G. 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