Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design

Autor(es)

Lu, Stephen ^[1] ; Ascencio, Mariano E. ^[2] ; Torquato, Ricardo J. S. ^[1] ; Florin-Christensen, Monica ^[2] ; Tanaka, Aparecida S. ^{[1, 3]}

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo UNIFESP, Escola Paulista Med, Dept Biochem, Rua 3 Maio 100, BR-04044020 Sao Paulo, SP - Brazil

^[2] INTA Castelar, Ctr Invest Ciencias Ve & Agron CICVyA, Inst Patobiol Vet, Los Reseros & Nicolas Repetto S-N, RA-1686 Hurlingham - Argentina

^[3] Inst Nacl Ciencia & Tecnol Entomol INCT EM, Rio De Janeiro - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Biochimie; v. 179, p. 127-134, DEC 2020.
• Citações Web of Science: 0

Resumo

C1A cysteine peptidases have been shown to play an important role during apicomplexan invasion and egress of host red blood cells (RBCs) and therefore have been exploited as targets for drug development, in which peptidase specificity is deterministic. Babesia bovis genome is currently available and from the 17 putative cysteine peptidases annotated four belong to the C1A subfamily. In this study, we describe the biochemical characterization of a C1A cysteine peptidase, named here BbCp (B. bovis cysteine peptidase) and evaluate its possible participation in the parasite asexual cycle in host RBCs. The recombinant protein was obtained in bacterial inclusion bodies and after a refolding process, presented typical kinetic features of the cysteine peptidase family, enhanced activity in the presence of a reducing agent, optimum pH between 6.5 and 7.0 and was inhibited by cystatins from R. microplus. Moreover, rBbCp substrate specificity evaluation using a peptide phage display library showed a preference for Val > Leu > Phe. Finally, antibodies anti-rBbCp were able to interfere with B. bovis growth in vitro, which highlights the BbCp as a potential target for drug design. (C) 2020 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved. (AU)

• Processo FAPESP: 15/09268-1 - Caracterização de duas proteases do parasita Babesia bovis: importância na infecção de seus hospedeiros.
• Beneficiário: Stephen Lu
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 17/25609-9 - Caracterização funcional de duas proteases de Babesia bovis e sua importância durante a infecção
• Beneficiário: Stephen Lu
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado
• Processo FAPESP: 12/03657-8 - Inibidores e proteases de ectoparasitas: relação de estrutura-função e identificação do papel dessas moléculas na interação de vetores de doenças e seus agentes etiológicos
• Beneficiário: Aparecida Sadae Tanaka
• Modalidade de apoio: Auxílio à Pesquisa - Temático