Identification and characterization of Harobin, a novel fibrino(geno)lytic serine protease from a sea snake (Lapemis hardwickii)

Abstract

A gene encoding a novel serine protease designated as Harobin is cloned and identified from a sea snake venom gland bacteriophage T7 library. It has 265 amino acids and shares 50-70% similarity to terrestrial snake serine proteases. In addition to the 12 conservative Cys, it has three more Cys residues that may contribute to its higher enzymatic stability. Harobin is expressed in Pichia pastoris and purified.Recombinant Harobin exhibits an amidolytic activity, and specifically degrades Aa, Bb-chain of fibrinogen. It functions as a defibrase both in vitro and in vivo, and reduces thrombosis. Harobin prolongs the coagulation time and the bleeding time of mice and reduces the fibrinogen levels of rats as well. Meanwhile, intravenous injection of Harobin leads to the reduction of blood pressure in SHR rats. It results from the ability of Harobin that cleaves angiotensin I and release bradykinin from plasma kininogen in vitro and in vivo. These data suggest that Harobin is a novel defibrase and has a potential to be an agent for the therapy of thrombosis and hypertension.

Coralsnake Venomics: Analyses of Venom Gland Transcriptomes and Proteomes of Six Brazilian Taxa

“…Given the great similarity of the micrurine serine proteases presented here, the M. fulvius sequences may represent contaminating blood or tissue serine proteases. When the M. paraensis sequence was used as a query string, the most similar sequence was a trypsin-like serine protease from Hydrophis hardwickii venom [224]. Harobin is anti-coagulant, and has both fibrinolytic and fibrinogenolytic activities.…”

What killed Karl Patterson Schmidt? Combined venom gland transcriptomic, venomic and antivenomic analysis of the South African green tree snake (the boomslang), Dispholidus typus

“…Fibrin(ogen)olytic serine proteases have isolated from the venom of Philodryas olfersii [75]. D. typus venom serine proteinases exhibit 75–78% amino acid sequence identity with a number of proteolytic enzymes from colubroid ( P. olfersii Q09GK1 [76]) and elapid venoms, including the fibrin(ogen)olytic enzymes Q5MCS0 ( Hydrophis hardwickii ) [77] and A8QL56 ( Ophiophagus hannah ) [78].…”

Bio-prospecting of cuttle fish waste and cow dung for the production of fibrinolytic enzyme from Bacillus cereus IND5 in solid state fermentation

“…In recent years, microbial fibrinolytic enzymes have been considered cost effective among the other fibrinolytic enzymes. These enzymes have been extensively studied from insects (Ahn et al 2003), snake venom (He et al 2007), and marine organisms (Sumi et al 1992). Fermented foods namely, natto (Sumi et al 1987, 1990; Fujita et al.…”

“…1993, 1995), Chungkook-Jang (Kim et al 1996) and Tempeh (Sugimoto et al 2007) have also been the source of fibrinolytic enzyme isolation. Apart from therapeutic applications, fibrinolytic proteases continue to be attractive as their potent activity on blood clot (He et al 2007), Keratin (Bressollier et al 1999) and collagen (Itoi et al 2006) could have important industrial waste management and medical applications.…”