title: Boophilus microplus cathepsin L-like (BmCL1) cysteine protease: specificity study using a peptide phage display library.
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.
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, Rua Três de Maio, 100, 04044-020 São Paulo, SP, Brazil.
authors.forename: Renan O
authors.initials: RO
authors.lastname: Clara
authors.forename: Tatiane S
authors.initials: TS
authors.lastname: Soares
authors.forename: Ricardo J S
authors.initials: RJ
authors.lastname: Torquato
authors.forename: Cássia A
authors.initials: CA
authors.lastname: Lima
authors.forename: Renata O M
authors.initials: RO
authors.lastname: Watanabe
authors.forename: Nilana M T
authors.initials: NM
authors.lastname: Barros
authors.forename: Adriana K
authors.initials: AK
authors.lastname: Carmona
authors.forename: Aoi
authors.initials: A
authors.lastname: Masuda
authors.forename: Itabajara S
authors.initials: IS
authors.lastname: Vaz
authors.forename: Aparecida S
authors.initials: AS
authors.lastname: Tanaka
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D057056:Cysteine Proteases / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D048494:Rhipicephalus / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D019151:Peptide Library; D057056:Cysteine Proteases