In silico analysis and functional characterization of a leucine-rich repeat protein of Leptospira interrogans.

Gaspar, João P; Takahashi, Maria B; Teixeira, Aline F; Nascimento, Ana L T O

Gaspar, João P; Takahashi, Maria B; Teixeira, Aline F; Nascimento, Ana L T O.

Afiliación

Gaspar JP; Laboratório de Desenvolvimento de Vacinas, Instituto Butantan, Avenida Vital Brazil, São Paulo, SP, Brazil; Programa de Pós-Graduação Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, SP, Brazil.
Takahashi MB; Laboratório de Desenvolvimento de Vacinas, Instituto Butantan, Avenida Vital Brazil, São Paulo, SP, Brazil.
Teixeira AF; Laboratório de Desenvolvimento de Vacinas, Instituto Butantan, Avenida Vital Brazil, São Paulo, SP, Brazil.
Nascimento ALTO; Laboratório de Desenvolvimento de Vacinas, Instituto Butantan, Avenida Vital Brazil, São Paulo, SP, Brazil. Electronic address: ana.nascimento@butantan.gov.br.

Int J Med Microbiol ; 316: 151633, 2024 Sep.

Article en En | MEDLINE | ID: mdl-39232290

ABSTRACT

ABSTRACT

Pathogenic spirochetes of the genus Leptospira are the causative agent of leptospirosis, a widely disseminated zoonosis that affects humans and animals. The ability of leptospires to quickly cross host barriers causing infection is not yet fully understood. Thus, understanding the mechanisms of pathogenicity is important to combat leptospiral infection. Outer membrane proteins are interesting targets to study as they are able to interact with host molecules. Proteins containing leucine-rich repeat (LRR) domains are characterized by the presence of multiple regions containing leucine residues and they have putative functions related to host-pathogen interactions. Hence, the present study aimed to clone and express the recombinant protein encoded by the LIC11098 gene, an LRR protein of L. interrogans serovar Copenhageni. In silico analyses predicted that the target protein is conserved among pathogenic strains of Leptospira, having a signal peptide and multiple LRR domains. The DNA sequence encoding the LRR protein was cloned in frame into the pAE vector, expressed without mutations in Escherichia coli and purified by His-tag chromatography. Circular dichroism (CD) spectrum showed that the recombinant protein was predominantly composed of ß-sheets. A dose-dependent interaction was observed with cellular and plasma fibronectins, laminin and the complement system component C9, suggesting a possible role of the protein encoded by LIC11098 gene at the initial stages of infection.

Asunto(s)

Leptospira interrogans; Proteínas Repetidas Ricas en Leucina; Proteínas Recombinantes; Leptospira interrogans/genética; Leptospira interrogans/metabolismo; Proteínas Recombinantes/genética; Proteínas Recombinantes/metabolismo; Proteínas Recombinantes/química; Simulación por Computador; Escherichia coli/genética; Escherichia coli/metabolismo; Humanos; Clonación Molecular; Proteínas Bacterianas/genética; Proteínas Bacterianas/metabolismo; Proteínas Bacterianas/química; Leptospirosis/microbiología; Animales; Interacciones Huésped-Patógeno; Proteínas de la Membrana Bacteriana Externa/genética; Proteínas de la Membrana Bacteriana Externa/metabolismo; Proteínas de la Membrana Bacteriana Externa/química; Dicroismo Circular; Secuencia de Aminoácidos

Palabras clave

Adhesin; Host-pathogen interaction; Leptospira; Leptospirosis; Leucine rich repeats; Recombinant proteins

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Recombinantes / Proteínas Repetidas Ricas en Leucina / Leptospira interrogans Límite: Animals / Humans Idioma: En Revista: Int J Med Microbiol Asunto de la revista: MICROBIOLOGIA Año: 2024 Tipo del documento: Article Pais de publicación: Alemania

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google