Intermediate and C-terminal regions of leptospiral adhesin Lsa66 are responsible for binding with plasminogen and extracellular matrix components

Abstract

Leptospirosis, a worldwide zoonotic infection, is an important human and veterinary health problem. We have previously identified a leptospiral multipurpose adhesin, Lsa66, capable of binding extracellular matrix (ECM) components and plasminogen (PLG). In this work, we report the cloning, expression, purification and characterization of three fragments derived from the full-length Lsa66: N-terminal, intermediate and C-terminal regions. We employed Escherichia coli BL21-SI as expression cells. The recombinant fragments tagged with N-terminal His 6 were purified by metal-charged chromatography to major protein bands that were recognized by anti-His-tag mAbs. The recombinant fragments were evaluated for their capacity to attach to ECM components and to PLG. The intermediate region bound to laminin, plasma fibronectin and PLG. Laminin also bound to the C-terminal region. Antibodies in leptospirosis-positive serum samples recognized Lsa66, being the immune epitopes located at the N-terminal and intermediate fragments. The data confirm that Lsa66 is expressed during infection and that this protein might have a role in bacterial infection.

Interação de proteínas de membrana de <i>Leptospira</i> com vitronectina humana.

“…As análises estatísticas foram realizadas utilizando ANOVA. (BARBOSA et al, 2006;BREINER et al, 2009;CHOY et al, 2007;DOMINGOS et al, 2015;LIN et al, 2007LIN et al, , 2008LIN et al, , 2009LIN et al, , 2010LIN et al, , 2011OLIVEIRA et al, 2014;SOUZA, et al, 2012 (BYKOWSKI et al, 2007;HALLSTRÖM et al, 2010HALLSTRÖM et al, , 2013KRAICZY et al, 2013;SELING et al, 2010). Em suma, a identificação de proteínas envolvidas no processo de colonização é fundamental para o conhecimento da patogenicidade da bactéria e poderá contribuir para o desenvolvimento de estratégias terapêuticas e/ou preventivas que interfiram no processo de evasão imune de leptospiras patogênicas.…”