Abstract
Ticks are rich sources of serine protease inhibitors, particularly those that prevent blood clotting and inflammatory responses during blood feeding. The tick Rhipicephalus (Boophlus) annulatus is an important ectoparasite of cattle. The aims of this study were to characterize and purify the serine protease inhibitors present in R. (B.) annulatus larval extract. The inhibitors were characterized by means of one and two-dimensional reverse zymography, and purified using affinity chromatography on a trypsin-Sepharose column. The analysis on one and two-dimensional reverse zymography of the larval extract showed trypsin inhibitory activity at between 13 and 40 kDa. Through non-reducing SDS-PAGE and reverse zymography for proteins purified by trypsin-Sepharose affinity chromatography, some protein bands with molecular weights between 13 and 34 kDa were detected. Western blotting showed that five protein bands at 48, 70, 110, 130 and 250 kDa reacted positively with immune serum, whereas there was no positive reaction in the range of 13-40 kDa. Serine protease inhibitors from R. (B.) annulatus have anti-trypsin activity similar to inhibitors belonging to several other hard tick species, thus suggesting that these proteins may be useful as targets in anti-tick vaccines.
Highlights
Ticks are the most frequent ectoparasites that infest mammals, birds, reptiles and amphibians (SAUER et al, 1995)
The inhibitors were characterized by means of one and two-dimensional reverse zymography, and purified using affinity chromatography on a trypsin-Sepharose column
Through non-reducing SDS-PAGE and reverse zymography for proteins purified by trypsin-Sepharose affinity chromatography, some protein bands with molecular weights between 13 and 34 kDa were detected
Summary
Ticks are the most frequent ectoparasites that infest mammals, birds, reptiles and amphibians (SAUER et al, 1995). In Iran, Rhipicephalus annulatus (formerly Boophilus annulatus) is one of the most common ticks in cattle. This tick has been recorded in some regions of Europe and the Americas, such as; Portugal, Spain, Italy, Greece, Turkey and Mexico; it is common in northern Africa. Tick control is mainly based on acaricides. Because of the disadvantages of acaricides, which including pollution of the environment and food chain and acaricide resistance (PREVOT et al, 2007), alternative control methods such as vaccination are promising. Development of vaccines relies on obtaining more knowledge about tick proteins. Proteases and their inhibitors may be candidates for designing vaccines
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Schedule a callMore From: Revista brasileira de parasitologia veterinaria = Brazilian journal of veterinary parasitology : Orgao Oficial do Colegio Brasileiro de Parasitologia Veterinaria
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