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- Title
Surface proteomics and label-free quantification of Leptospira interrogans serovar Pomona.
- Authors
Techawiwattanaboon, Teerasit; Thaibankluay, Praparat; Kreangkaiwal, Chahya; Sathean-Anan-Kun, Suwitra; Khaenam, Prasong; Makjaroen, Jiradej; Pisitkun, Trairak; Patarakul, Kanitha
- Abstract
Leptospirosis is a re-emerging zoonosis with a global distribution. Surface-exposed outer membrane proteins (SE-OMPs) are crucial for bacterial–host 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 Leptospira interrogans serovar Pomona. Two complementary approaches, surface biotinylation and surface proteolytic shaving, followed by liquid chromatography tandem-mass spectrometry (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 βb-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. Author summary: Surface-exposed outer membrane proteins (SE-OMPs) are important components of pathogenic Leptospira mediating virulence and interacting with host cells and host environment. This study aimed to identify SE-OMPs of pathogenic L. interrogans serovar Pomona using surface biotinylation and proteinase K (proK) shaving methods comprehensively. Surface-protein enriched fractions obtained from intact leptospires were subsequently subjected to liquid chromatography tandem-mass spectrometry (LC-MS/MS) and their label-free quantitative profiles were analyzed by MaxQuant software. The surface biotinylation and proK shaving yielded a total of 1,001 and 238 proteins, respectively, and shared 220 proteins. Of these, 28 known SE-OMPs were identified by either method and 11 known SE-OMPs were overlapped. Moreover, 50 predicted SE-OMPs were detected in individual sample group and 18 were common in both groups. In the 20 most abundant proteins, there were six known SE-OMPs derived from both methods, including EF- Tu, LipL21, LipL41, LipL46, Loa22, and OmpL36, and one predicted SE-OMP, LipL71. McpA, OmpL1, OmpL32, OmpL36, SdhA, SppA, LIC10314, and LIC12615 were ranked in the 50 most abundant proteins. Furthermore, LIC10411 (LIP3228 ortholog in serovar Pomona) was predicted as a novel surface βb-OMP. The high-abundance SE-OMPs should be further investigated as novel vaccine candidates or diagnostic biomarkers for leptospirosis.
- Subjects
POMONA (Calif.); LEPTOSPIRA interrogans; PROTEOMICS; MEMBRANE proteins; LIQUID chromatography-mass spectrometry; LIQUID chromatography; VACCINE development
- Publication
PLoS Neglected Tropical Diseases, 2021, Vol 15, Issue 11, p1
- ISSN
1935-2727
- Publication type
Article
- DOI
10.1371/journal.pntd.0009983