Characterization of a virulence-modifying protein of Leptospira interrogans identified by shotgun phage display

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dc.contributorCentro Bioindustrialpt_BR
dc.contributorLab. Bacteriologiapt_BR
dc.contributor.authorFerreira, Fabiana Laurettipt_BR
dc.contributor.authorTeixeira, André Azevedo Reispt_BR
dc.contributor.authorGiordano, Ricardo Josépt_BR
dc.contributor.authorSilva, Josefa Bezerra dapt_BR
dc.contributor.authorAbreu, Patricia Antonia Estimapt_BR
dc.contributor.authorBarbosa, Angela Silvapt_BR
dc.contributor.authorAkamatsu, Milena Apetitopt_BR
dc.contributor.authorHo, Paulo Leept_BR
dc.identifier.citationFerreira FL, Teixeira AAR, Giordano RJ, Silva JB, Abreu PAE, Barbosa AS, et al. Characterization of a virulence-modifying protein of Leptospira interrogans identified by shotgun phage display. Front Microbiol. 2022 Nov; 13:1051698. doi:10.3389/fmicb.2022.1051698.pt_BR
dc.description.abstractPathogenic species of Leptospira are etiologic agents of leptospirosis, an emerging zoonotic disease of worldwide extent and endemic in tropical regions. The growing number of identified leptospiral species sheds light to their genetic diversity and unique virulence mechanisms, many of them still remain unknown. Toxins and adhesins are important virulence factors in several pathogens, constituting promising antigens for the development of vaccines with cross-protection and long-lasting effect against leptospirosis. For this aim, we used the shotgun phage display technique to unravel new proteins with adhesive properties. A shotgun library was constructed using fragmented genomic DNA from Leptospira interrogans serovar Copenhageni strain Fiocruz L1-130 and pG8SAET phagemid vector. Selection of phages bearing new possible cell-binding antigens was performed against VERO cells, using BRASIL biopanning methodology. Analysis of selected clones revealed the hypothetical protein LIC10778, a potentially exposed virulence factor that belongs to the virulence-modifying (VM) protein family (PF07598), composed of 13 members in the leptospiral strain Fiocruz L1-130. Prediction of LIC10778 tertiary structure indicates that the protein contains a cellular-binding domain (N-terminal portion) and an unknown domain of no assigned activity (C-terminal portion). The predicted N-terminal domain shared structural similarities with the cell-binding and internalization domain of toxins like Ricin and Abrin, as well as to the Community-Acquired Respiratory Distress Syndrome (CARDS) toxin in Mycoplasma pneumoniae. Interestingly, recombinant portions of the N-terminal region of LIC10778 protein showed binding to laminin, collagens I and IV, vitronectin, and plasma and cell fibronectins using overlay blotting technique, especially regarding the binding site identified by phage display. These data validate our preliminary phage display biopanning and support the predicted three-dimensional models of LIC10778 protein and other members of PF07598 protein family, confirming the identification of the N-terminal cell-binding domains that are similar to ricin-like toxins. Moreover, fluorescent fused proteins also confirmed that N-terminal region of LIC10778 is capable of binding to VERO and A549 cell lines, further highlighting its virulence role during host-pathogen interaction in leptospirosis probably mediated by its C-terminal domain. Indeed, recent results in the literature confirmed this assumption by demonstrating the cytotoxicity of a closely related PF07598 member.pt_BR
dc.description.sponsorship(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulopt_BR
dc.description.sponsorship(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológicopt_BR
dc.relation.ispartofFrontiers in Microbiologypt_BR
dc.rightsOpen accesspt_BR
dc.titleCharacterization of a virulence-modifying protein of Leptospira interrogans identified by shotgun phage displaypt_BR
dc.rights.licenseCC BYpt_BR
dc.contributor.external(USP) Universidade de São Paulopt_BR
dc.subject.keywordphage displaypt_BR
dc.subject.keywordbinding domainspt_BR
dc.subject.keywordprotein structure predictionpt_BR
dc.relation.ispartofabbreviatedFront Microbiolpt_BR
dc.identifier.citationabntv. 13, 1051698, nov. 2022pt_BR
dc.identifier.citationvancouver2022 Nov; 13:1051698pt_BR
dc.contributor.butantanFerreira, Fabiana Lauretti|:Técnico|:Centro Bioindustrial|:Primeiroautorpt_BR
dc.contributor.butantanSilva, Josefa Bezerra da|:Pesquisador|:Lab. Bacteriologiapt_BR
dc.contributor.butantanAbreu, Patricia Antonia Estima|:Pesquisador|:Lab. Bacteriologiapt_BR
dc.contributor.butantanBarbosa, Angela Silva|:Pesquisador|:Lab. Bacteriologiapt_BR
dc.contributor.butantanAkamatsu, Milena Apetito|:Outros|:Centro Bioindustrialpt_BR
dc.contributor.butantanHo, Paulo Lee|:Pesquisador|:Centro Bioindustrial|:Autor de correspondênciapt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦2013/15155–0pt_BR
dc.sponsorship.butantan(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológico¦¦164209/2015–8pt_BR
dc.sponsorship.butantan(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológico¦¦305430/2019–0pt_BR
dc.sponsorship.butantan(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológico¦¦148645/2013-5pt_BR
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