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

Author(s)

Clara, Renan O. ^[1] ; Soares, Tatiane S. ^[1] ; Torquato, Ricardo J. S. ^[1] ; Lima, Cassia A. ^[1] ; Watanabe, Renata O. M. ^[1] ; Barros, Nilana M. T. ^[2] ; Carmona, Adriana K. ^[2] ; Masuda, Aoi ^[3] ; Vaz Junior, Itabajara S. ^[3] ; Tanaka, Aparecida S. ^[1]

Affiliation

^[1] Univ Fed Sao Paulo, Dept Bioquim, BR-04044020 Sao Paulo - Brazil

^[2] Univ Fed Sao Paulo, Dept Biofis, BR-04044020 Sao Paulo - Brazil

^[3] Univ Fed Rio Grande do Sul, Ctr Biotecnol Estado Rio Grande Sul, Porto Alegre, RS - Brazil

• Document type: Journal article
• Source: Veterinary Parasitology; v. 181, n. 2-4, p. 291-300, SEP 27 2011.
• Web of Science Citations: 13

Abstract

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. (AU)

• FAPESP's process: 05/03514-9 - Studies of the physiological function and biotechnological potential of protease inhibitors and anti-hemostatics in hematophagous arthropods
• Grantee: Aparecida Sadae Tanaka
• Support Opportunities: Research Projects - Thematic Grants