title: A newly identified leptospiral adhesin mediates attachment to laminin.
abstract: Pathogenic leptospires have the ability to survive and disseminate to multiple organs after penetrating the host. Several pathogens, including spirochetes, have been shown to express surface proteins that interact with the extracellular matrix (ECM). This adhesin-mediated binding process seems to be a crucial step in the colonization of host tissues. This study examined the interaction of putative leptospiral outer membrane proteins with laminin, collagen type I, collagen type IV, cellular fibronectin, and plasma fibronectin. Six predicted coding sequences selected from the Leptospira interrogans serovar Copenhageni genome were cloned, and proteins were expressed, purified by metal affinity chromatography, and characterized by circular dichroism spectroscopy. Their capacity to mediate attachment to ECM components was evaluated by binding assays. We have identified a leptospiral protein encoded by LIC12906, named Lsa24 (leptospiral surface adhesin; 24 kDa) that binds strongly to laminin. Attachment of Lsa24 to laminin was specific, dose dependent, and saturable. Laminin oxidation by sodium metaperiodate reduced the protein-laminin interaction in a concentration-dependent manner, indicating that laminin sugar moieties are crucial for this interaction. Triton X-114-solubilized extract of L. interrogans and phase partitioning showed that Lsa24 was exclusively in the detergent phase, indicating that it is a component of the leptospiral membrane. Moreover, Lsa24 partially inhibited leptospiral adherence to immobilized laminin. This newly identified membrane protein may play a role in mediating adhesion of L. interrogans to the host. To our knowledge, this is the first leptospiral adhesin with laminin-binding properties reported to date.
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900, São Paulo, Brazil.
authors.forename: Angela S
authors.initials: AS
authors.lastname: Barbosa
authors.forename: Patricia A E
authors.initials: PA
authors.lastname: Abreu
authors.forename: Fernanda O
authors.initials: FO
authors.lastname: Neves
authors.forename: Marina V
authors.initials: MV
authors.lastname: Atzingen
authors.forename: Mônica M
authors.initials: MM
authors.lastname: Watanabe
authors.forename: Mônica L
authors.initials: ML
authors.lastname: Vieira
authors.forename: Zenaide M
authors.initials: ZM
authors.lastname: Morais
authors.forename: Sílvio A
authors.initials: SA
authors.lastname: Vasconcellos
authors.forename: Ana L T O
authors.initials: AL
authors.lastname: Nascimento
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology*; D001422:Bacterial Adhesion / Q000502:physiology*; D007797:Laminin / Q000378:metabolism*; D007921:Leptospira interrogans / Q000235:genetics / Q000502:physiology*; D011485:Protein Binding / Q000276:immunology
chemical_list: D018829:Adhesins, Bacterial; D007797:Laminin