Cloning and functional expression of a Boophilus microplus cathepsin L-like enzyme

Gaby Renard, José F. Garcia, Felipe C. Cardoso, Marc F. Richter, Judy A. Sakanari, Luiz S. Ozaki, Carlos Termignoni, Aoi Masuda

Research output: Contribution to journalArticle

Abstract

A cysteine proteinase gene homologous to cathepsins L genes was isolated from a B. microplus cDNA library. The precursor protein deduced from the nucleotide sequence contains 332 amino acid residues consisting of a signal sequence (pre-region), a pro-region and a mature proteinase. The DNA fragment coding for the proenzyme was cloned and expressed using the E. coli expression vector pMAL-p. The recombinant protein (MBP+PROCP) once activated is able to hydrolyze synthetic substrates as well asiprotein substrates like hemoglobin, vitellin and gelatin. Its optimal enzymatic activity on both fluorogenic and protein substrates was found to occur at an acidic pH. Expression of the proteinase gene was tested by RT-PCR with tick larvae RNA. Detection of amplified sequences indicates that the gene is expressed at this stage of the tick life cycle and the molecule is therefore potentially a target for chemotherapy or an immunogen in a vaccine. (C) 2000 Elsevier Science Ltd.

Original languageEnglish (US)
Pages (from-to)1017-1026
Number of pages10
JournalInsect Biochemistry and Molecular Biology
Volume30
Issue number11
DOIs
StatePublished - Oct 19 2000
Externally publishedYes

Keywords

  • Boophilus microplus
  • Cathepsin L
  • Cysteine proteinase
  • Gene expression
  • cDNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Insect Science

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    Renard, G., Garcia, J. F., Cardoso, F. C., Richter, M. F., Sakanari, J. A., Ozaki, L. S., Termignoni, C., & Masuda, A. (2000). Cloning and functional expression of a Boophilus microplus cathepsin L-like enzyme. Insect Biochemistry and Molecular Biology, 30(11), 1017-1026. https://doi.org/10.1016/S0965-1748(00)00070-9