title: Evaluation of the elastinolytic activity and protective effect of Leptallo I, a protein composed by metalloprotease and FA5/8C domains, from Leptospira interrogans Copenhageni.
abstract: Leptospirosis is a re-emergent zoonosis, caused by pathogenic spirochetes from the genus Lepstospira. To date, there is no protein described to be involved in leptospiral hemorrhagic manifestations, although several proteases have been reported for other bacterial infections. In this study we identified 12 putative metalloproteases from the genome of Leptospira interrogans, and characterized for the first time a putative metalloprotease, here named Leptallo I, as a potential Zn(2+) dependent glycylglycine protease belonging to the M23 metalloendopeptidase family. The native protein was detected in extracts from several pathogenic Leptospira species and further shown to be secreted to the culture medium. We expressed the recombinant protein and its C-terminal fragment containing the metalloprotease domain, and both presented regular secondary structures. The sera of humans with leptospirosis were able to recognize rLeptallo I, indicating that the native protein is expressed and presented to the immune system during infection. The recombinant proteins displayed a significant, though relatively low, elastinolytic activity, and the challenge of hamsters immunized with rLeptallo I conferred 33% protection, suggesting a significant importance of this protein in the pathogenesis. The elastinolytic activity may be important for leptospires-host interaction, because elastin constitutes a significant proportion of total lung and blood vessel proteins.
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, 05503-900 São Paulo, SP, Brazil.
authors.forename: V L
authors.initials: VL
authors.lastname: Hashimoto
authors.forename: P A E
authors.initials: PA
authors.lastname: Abreu
authors.forename: E
authors.initials: E
authors.lastname: Carvalho
authors.forename: A P
authors.initials: AP
authors.lastname: Gonçales
authors.forename: Z M
authors.initials: ZM
authors.lastname: Morais
authors.forename: S A
authors.initials: SA
authors.lastname: Vasconcellos
authors.forename: E C
authors.initials: EC
authors.lastname: Romero
authors.forename: P L
authors.initials: PL
authors.lastname: Ho
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D006224:Cricetinae; D004549:Elastin / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007921:Leptospira interrogans / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D007922:Leptospirosis / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D008297:Male; D008647:Mesocricetus; D045726:Metalloproteases / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010196:Pancreatic Elastase / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D004549:Elastin; D045726:Metalloproteases; D010196:Pancreatic Elastase
keywords: Leptospira; Leptospiral pulmonary hemorrhagic syndrome; Leptospirosis; Metalloprotease