Purification and biological effects of a C-type lectin isolated from Bothrops moojeni

Abstract

Snake venom proteins from the C-type lectin family have very distinct biological activities despite their highly conserved primary structure, which is homologous to the carbohydrate recognition region of true C-type lectins. We purified a lectin-like protein (BmLec) from Bothrops moojeni venom and investigated its effect on platelet aggregation, insulin secretion, antibacterial activity, and isolated kidney cells. The BmLec was purified using two chromatographic steps: affinity chromatography and reverse phase high performance liquid chromatography (HPLC). BmLec showed a dose-dependent platelet aggregation and significantly decreased the bacterial growth rate in approximately 15%. During scanning electron microscopy, the profile of Xanthomonas axonopodis pv. passiflorae treated with lectin disclosed a high vesiculation and membrane rupture. BmLec induced a strong and significant increase in insulin secretion at 2.8 and 16.7 mM glucose concentrations, and this effect was seen in the presence of EGTA in both experiments. BmLec (10 µg/mL) increased the perfusion pressure, renal vascular resistance and urinary flow. The glomerular filtration rate and percentages of sodium, potassium and chloride tubular transport were reduced at 60 minutes of perfusion. Renal alterations caused by BmLec were completely inhibited by indomethacin in all evaluated parameters. In conclusion, the C-type lectin isolated from Bothrops moojeni affected platelet aggregation, insulin secretion, antibacterial activity and isolated kidney function

Purification of a lectin with antibacterial activity from Bothrops leucurus snake venom

“…Glycoconjugates present on bacterial cell surfaces, such as peptidoglycans, lipopolysaccharides and teichoic acids, constitute potential lectin targets (Lee et al, 1998;Santi-Gadelha et al, 2006). Recently, it was reported that snake venom lectins are able to inhibit growth of phytopathogenic bacteria (Rádis-Baptista et al, 2006;Barbosa et al, 2010); however, the interactions between snake venom lectins and human pathogenic bacteria have not been studied.…”

In vitroandin vivogenotoxic evaluation ofBothrops moojenisnake venom

“…For example, Barbosa et al (2010) reported the in vitro antibacterial and antiplatelet aggregation activities, the insulin secretion stimulation, and the in vivo improvement of kidney function for a lectin-like protein isolated from BMV. Morais et al (2012) showed the anticoagulant and fibrinogenolytic activities of moojenin, a metalloproteinase extracted from BMV.…”

Snake venom galactoside-binding lectins: a structural and functional overview

“…All SVgalLs are homodimeric proteins composed of disulfide-linked monomers presenting molecular mass varying from 14 to 16.2 kDa. The primary structures of 12 SVgalLs were determined, and presented from 134 to 136 amino acid residues as described for the following lectins: RSL—rattlesnake lectin from Crotalus atrox [ 23 ], ApL— Agkistrodon piscivorus piscivorus lectin [ 15 ], BaL— Bitis arietans lectin [ 13 ], CrL— Crotalus ruber lectin [ 18 ], BiL— Bothrops insularis lectin [ 16 ], BmLec— Bothrops moojeni lectin [ 24 ], BpalL— Bothrops pauloensis lectin [ 21 ], BJcuL— Bothrops jararacussu lectin [ 25 ], BpirL— Bothrops pirajai lectin [ 17 ], LmL— Lachesis muta stenophrys lectin [ 26 ], ToL— Trimeresurus okinavensis lectin [ 27 ] and TsL— Trimeresurus stejnegeri lectin [ 28 ]. Amino acid sequence analysis among the referenced SVgalLs using BLAST searching tool [ 29 ] showed an identity degree from 82 to 97 % among them, indicating a high primary structural similarity within these lectins.…”

“…Carbohydrate recognition domain (CRD) is indicated by the blue box. Primary structure according to lectins: RSL [GI: 126130], ApL [GI: 510120659], BaL [GI: 34922645], CrL [GI: 118572769], BiL [GI: 82126834], BmLec [ 24 ], BpalL [GI: 527504051], BJcuL [ 25 ], BpirL [GI: 510120660], LmL [GI: 1881829], ToL [ 27 ] and TsL [GI: 7674107] …”