title: Assessment and partial purification of serine protease inhibitors from Rhipicephalus (Boophilus) annulatus larvae.
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.
authors: Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran ? UT, Tehran, Iran.
authors: Sedigheh
authors: S
authors: Nabian
authors: Rastegar Reference Laboratory, Faculty of Veterinary Medicine, University of Tehran ? UT, Tehran, Iran.
authors: Mohammad
authors: M
authors: Taheri
authors: Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran ? UT, Tehran, Iran.
authors: Mohammad Mehdi
authors: MM
authors: Ranjbar
authors: Institute of Parasitology, Department of Pathobiology, University of Veterinary Medicine Vienna ? Vetmeduni Vienna, Vienna, Austria.
authors: Alireza
authors: A
authors: Sazmand
authors: Rastegar Reference Laboratory, Faculty of Veterinary Medicine, University of Tehran ? UT, Tehran, Iran.
authors: Parastou
authors: P
authors: Youssefy
authors: Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran ? UT, Tehran, Iran.
authors: Gholam Reza
authors: GR
authors: Nazaralipour
mesh_terms: D000818:Animals; D007814:Larva / Q000737:chemistry; D011506:Proteins; D048494:Rhipicephalus / Q000737:chemistry*; D015842:Serine Proteinase Inhibitors / Q000302:isolation & purification*
chemical_list: D011506:Proteins; D015842:Serine Proteinase Inhibitors