title: Novel Leptospira interrogans protein Lsa32 is expressed during infection and binds laminin and plasminogen.
abstract: Pathogenic Leptospira is the aetiological agent of leptospirosis, a life-threatening disease of human and veterinary concern. The quest for novel antigens that could mediate host-pathogen interactions is being pursued. Owing to their location, these antigens have the potential to elicit numerous activities, including immune response and adhesion. This study focuses on a hypothetical protein of Leptospira, encoded by the gene LIC11089, and its three derived fragments: the N-terminal, intermediate and C terminus regions. The gene coding for the full-length protein and fragments was cloned and expressed in Escherichia coli BL21(SI) strain by using the expression vector pAE. The recombinant protein and fragments tagged with hexahistidine at the N terminus were purified by metal affinity chromatography. The leptospiral full-length protein, named Lsa32 (leptospiral surface adhesin, 32 kDa), adheres to laminin, with the C terminus region being responsible for this interaction. Lsa32 binds to plasminogen in a dose-dependent fashion, generating plasmin when an activator is provided. Moreover, antibodies present in leptospirosis serum samples were able to recognize Lsa32. Lsa32 is most likely a new surface protein of Leptospira, as revealed by proteinase K susceptibility. Altogether, our data suggest that this multifaceted protein is expressed during infection and may play a role in host-L. interrogans interactions.
authors: Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil Programa Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, USP, Avenida Professor Lineu Prestes, 1730, 05508-900, São Paulo, SP, Brazil.
authors: Renan F
authors: RF
authors: Domingos
authors: Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil Programa Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, USP, Avenida Professor Lineu Prestes, 1730, 05508-900, São Paulo, SP, Brazil.
authors: Luis G
authors: LG
authors: Fernandes
authors: Divisão de Biologia Medica, Instituto Adolfo Lutz, São Paulo, SP, Brazil.
authors: Eliete C
authors: EC
authors: Romero
authors: Laboratório de Zoonoses Bacterianas do VPS, Faculdade de Medicina Veterinária e Zootecnia, USP, Avenida Professor Dr Orlando Marques de Paiva, 87, 05508-270, São Paulo, SP, Brazil.
authors: Zenaide M
authors: ZM
authors: de Morais
authors: Laboratório de Zoonoses Bacterianas do VPS, Faculdade de Medicina Veterinária e Zootecnia, USP, Avenida Professor Dr Orlando Marques de Paiva, 87, 05508-270, São Paulo, SP, Brazil.
authors: Silvio A
authors: SA
authors: Vasconcellos
authors: Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil Programa Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, USP, Avenida Professor Lineu Prestes, 1730, 05508-900, São Paulo, SP, Brazil ana.nascimento@butantan.gov.br.
authors: Ana L T O
authors: AL
authors: Nascimento
mesh_terms: D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D019295:Computational Biology; D005109:Extracellular Matrix; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D007797:Laminin / Q000378:metabolism*; D007921:Leptospira interrogans / Q000235:genetics* / Q000378:metabolism*; D007922:Leptospirosis / Q000382:microbiology*; D010958:Plasminogen / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D007797:Laminin; D010958:Plasminogen