title: Lsa63, a newly identified surface protein of Leptospira interrogans binds laminin and collagen IV.
abstract: Leptospira interrogans is the etiological agent of leptospirosis, a zoonotic disease that affects populations worldwide. We have identified in proteomic studies a protein that is encoded by the gene LIC10314 and expressed in virulent strain of L. interrogans serovar Pomona. This protein was predicted to be surface exposed by PSORT program and contains a p83/100 domain identified by BLAST analysis that is conserved in protein antigens of several strains of Borrelia and Treponema spp. The proteins containing this domain have been claimed antigen candidates for serodiagnosis of Lyme borreliosis. Thus, we have cloned the LIC10314 and expressed the protein in Escherichia coli BL21-SI strain by using the expression vector pAE. The recombinant protein tagged with N-terminal hexahistidine was purified by metal-charged chromatography and characterized by circular dichroism spectroscopy. This protein is conserved among several species of pathogenic Leptospira and absent in the saprophytic strain L. biflexa. We confirm by liquid-phase immunofluorescence assays with living organisms that this protein is most likely a new surface leptospiral protein. The ability of the protein to mediate attachment to ECM components was evaluated by binding assays. The leptospiral protein encoded by LIC10314, named Lsa63 (Leptospiral surface adhesin of 63kDa), binds strongly to laminin and collagen IV in a dose-dependent and saturable fashion. In addition, Lsa63 is probably expressed during infection since it was recognized by antibodies of serum samples of confirmed-leptospirosis patients in convalescent phase of the disease. Altogether, the data suggests that this novel identified surface protein may be involved in leptospiral pathogenesis.
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900 São Paulo, SP, Brazil.
authors.forename: Monica L
authors.initials: ML
authors.lastname: Vieira
authors.forename: Zenaide M
authors.initials: ZM
authors.lastname: de Morais
authors.forename: Amane P
authors.initials: AP
authors.lastname: Gonçales
authors.forename: Eliete C
authors.initials: EC
authors.lastname: Romero
authors.forename: Silvio 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*; D001422:Bacterial Adhesion / Q000235:genetics; D001483:Base Sequence; D024141:Collagen Type IV / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D007921:Leptospira interrogans / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D007922:Leptospirosis / Q000276:immunology; D008969:Molecular Sequence Data
chemical_list: D018829:Adhesins, Bacterial; D024141:Collagen Type IV; D007797:Laminin