A newly identified protein of Leptospira interrogans mediates binding to laminin.
Pathogenic Leptospira is the aetiological agent of leptospirosis, a life-threatening disease that affects populations worldwide. The search for novel antigens that could be relevant in host-pathogen interactions is being pursued. These antigens have the potential to elicit several activities, including adhesion. This study focused on a hypothetical predicted lipoprotein of Leptospira, encoded by the gene LIC12895, thought to mediate attachment to extracellular matrix (ECM) components. The gene was cloned and expressed in Escherichia coli BL21 Star (DE3)pLys 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. The capacity of the protein to mediate attachment to ECM components was evaluated by binding assays. The leptospiral protein encoded by LIC12895, named Lsa27 (leptospiral surface adhesin, 27 kDa), bound strongly to laminin in a dose-dependent and saturable fashion. Moreover, Lsa27 was recognized by antibodies from serum samples of confirmed leptospirosis specimens in both the initial and the convalescent phases of the disease. Lsa27 is most likely a surface protein of Leptospira as revealed in liquid-phase immunofluorescence assays with living organisms. Taken together, these data indicate that this newly identified membrane protein is expressed during natural infection and may play a role in mediating adhesion of L. interrogans to its host.
Longhi
Mariana T
MT
Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil 1500, 05503-900 São Paulo, SP, Brazil.
Oliveira
Tatiane R
TR
Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil 1500, 05503-900 São Paulo, SP, Brazil.
Romero
Eliete C
EC
Divisão de Biologia Medica, Instituto Adolfo Lutz, Sao Paulo, SP, Brazil.
Gonçales
Amane P
AP
Laboratório de Zoonoses Bacterianas do VPS, Faculdade de Medicina Veterinária e Zootecnia, USP, Avenida Prof. Dr Orlando Marques de Paiva 87, 05508-270 São Paulo, SP, Brazil.
de Morais
Zenaide M
ZM
Laboratório de Zoonoses Bacterianas do VPS, Faculdade de Medicina Veterinária e Zootecnia, USP, Avenida Prof. Dr Orlando Marques de Paiva 87, 05508-270 São Paulo, SP, Brazil.
Vasconcellos
Silvio A
SA
Laboratório de Zoonoses Bacterianas do VPS, Faculdade de Medicina Veterinária e Zootecnia, USP, Avenida Prof. Dr Orlando Marques de Paiva 87, 05508-270 São Paulo, SP, Brazil.
Nascimento
Ana L T O
ALTO
Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, USP, Avenida Prof. Lineu Prestes 1730, 05508-900 São Paulo, SP, Brazil.|Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil 1500, 05503-900 São Paulo, SP, Brazil.
D018829:Adhesins, Bacterial / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000235:genetics / Q000378:metabolism; D001422:Bacterial Adhesion / Q000235:genetics / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D005798:Genes, Bacterial; D054884:Host-Pathogen Interactions / Q000276:immunology / Q000502:physiology; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D007921:Leptospira interrogans / Q000235:genetics* / Q000276:immunology / Q000378:metabolism* / Q000472:pathogenicity; D007922:Leptospirosis / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D017931:DNA Primers; D007797:Laminin; D008074:Lipoproteins; D011994:Recombinant Proteins