Boophilus microplus cathepsin L-like (BmCL1) cysteine protease: Specificity study using a peptide phage display library

• Autor(a) principal: Clara, Renan Orsati [UNIFESP]
• Data de Publicação: 2011
• Outros Autores: Soares, Tatiane Sanches [UNIFESP], Torquato, Ricardo Jose Soares [UNIFESP], Lima, Cassia Arantes de [UNIFESP], Watanabe, Renata Midori Okuta [UNIFESP], Barros, Nilana Meza Tenório de [UNIFESP], Carmona, Adriana Karaoglanovic [UNIFESP], Masuda, Aoi, Vaz Junior, Itabajara S., Tanaka, Aparecida Sadae [UNIFESP]
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNIFESP
• Texto Completo: http://repositorio.unifesp.br/handle/11600/34058; http://dx.doi.org/10.1016/j.vetpar.2011.04.003
• topic: Cysteine proteases; Rhipicephalus (Boophilus) microplus; Protein expression; Enzyme kinetic; Phage display library

The tick Rhipicephalus (Boophilus) microplus is one of the most important bovine ectoparasites, a disease vector responsible for losses in meat and milk productions. A cysteine protease similar to cathepsin L, named BmCL1, was previously identified in R. microplus gut, suggesting a role of the enzyme in meal digestion. in this work. BmCL1 was successfully expressed in Pichia pastoris system, yielding 54.8 mg/L of culture and its activity was analyzed by synthetic substrates and against a R. microplus cysteine protease inhibitor, Bmcystatin. After rBmCl1 biochemical characterization it was used in a selection of a peptide phage library to determine rBmCL1 substrate preference. Obtained sequenced clones showed that rBmCL1 has preference for Leu or Arg at P(1) position. the preference for Leu at position P(1) and the activation of BmCL1 after a Leu amino acid residue suggest possible self activation. (C) 2011 Elsevier B.V. All rights reserved.