Expression and immunolocalization of a Boophilus microplus cathepsin L-like enzyme.
Efforts are being undertaken to control tick infestations that cause important economic losses. A cathepsin L-like endopeptidase of Boophilus microplus was expressed in Escherichia coli; the recombinant enzyme was capable of hydrolysing gelatin, tick vitellin and bovine haemoglobin. In this paper we focus on the expression and local of synthesis of this enzyme in the tick. RT-PCR experiments showed that this endopeptidase is transcribed in the gut of partially engorged tick females. In immunoblotting, polyclonal antibodies against the recombinant enzyme reacted with proteins of larvae older than 5 days, of fully and partially engorged female gut. In immunolocalization experiments the enzyme was localized in probable secretory cells of the gut. Based on our findings we postulate that BmCL1 may be involved in haemoglobin degradation in the B. microplus gut. This enzyme may be used as target for the control of this parasite.
Renard
Gaby
G
Centro de Biotecnologia do Estado do Rio Grande do Sul, Departamento de Bioquímica and Departamento de Biologia Molecular and Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil. gaby@dna.cbiot.ufrgs.br
Lara
Flávio Alves
FA
de Cardoso
Felipe Cardoso
FC
Miguens
Flávio Costa
FC
Dansa-Petretski
Marílvia
M
Termignoni
Carlos
C
Masuda
Aoi
A
D000818:Animals; D015153:Blotting, Western; D056668:Cathepsin L; D002403:Cathepsins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D008854:Microscopy, Electron; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013987:Ticks / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure
D011994:Recombinant Proteins; D012313:RNA; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D056668:Cathepsin L