Purification and characterization of a metalloproteinase, Porthidin-1, from the venom of Lansberg’s hog-nosed pitvipers (Porthidium lansbergii hutmanni)

Abstract

Porthidium lansbergii hutmanni is a small pit viper found on Margarita Island, Venezuela. Local tissue damage is one of the most obvious characteristics of P. l. hutmanni envenomation, which can lead to diverse pathological effects, such as hemorrhage, edema, blistering, necrosis, lymphatic vessel damage and degradation of extracellular matrix. Metalloproteinases are one of the major components in venoms responsible for these effects. To date, very little is known or has been reported on P. l. hutmanni venom. Crude P. l. hutmanni venom had a LD50 of 2.5 mg/kg and was considered very hemorrhagic (minimal hemorrhagic dose [MHD]: 0.98 μg) when compared to other hemorrhagic (Bothrops) venoms in Venezuela. Crude P. l. hutmanni venom also inhibited ADP-induced platelet aggregation. A metalloproteinase, Porthidin-1, from this venom was isolated by three chromatography steps (Sephadex G100, Superose 12 HR10/30 and Bioscale Q2). Porthidin-1 falls in the SVMP P-I class having a molecular weight of 23 kDa, verified by both SDS-PAGE and mass spectrometry. High-resolution mass spectrometry and a database search identified a peptide from Porthidin-1 (YNGDLDK) belonging to the SVMP family of proteins. Porthidin-1 contained hemorrhagic, fibrino(geno)lytic, caseinolytic and gelatinolytic activities, and these activities were capable of being neutralized by metalloproteinase inhibitors but not serine proteinase inhibitors. The peptide YNGDLDK shared similarities with five venom proteins with a BLAST e-value of <1. This work details the biochemical and pathophysiological effects that can result from envenomations, and highlights the importance and significance for characterizing unknown or poorly documented venoms from different geographical regions.

Exploring the biochemical, haemostatic and toxinological aspects of mapanare dry-tail (Porthidium lansbergii rozei) snake´s venom causing human accidents in Eastern Venezuela

“…In the haemorrhagic activity evaluation, which is one of the characteristic effects of Porthidium venoms (7,8,26,30,31), it was observed that the P.l.rozei venom exhibits a 13.87 ± 3.2 μg / mouse MHD, which is high when compared with previous reports for this genus (Table II), in which P. l. hutmanni (8) reported a 0.98 μg/mouse MHD. Porthidium nasutum from Costa Rica (32) obtained a 0.5 μg/mouse MHD, while Gutiérrez et al (33) reported a 1.6 μg/mouse MHD.…”

“…A number of reports in the literature have documented different levels of variation in the composition of Porthidium venom (7,8). These differentiations may be related to phylogenetic or taxonomic discrepancies, age, size, geographic location, diet, seasonal variations, variations associated with sex, among others, all these lead to divergences in the quantity and quality of the venoms.…”

PhTX-II a Basic Myotoxic Phospholipase A2 from Porthidium hyoprora Snake Venom, Pharmacological Characterization and Amino Acid Sequence by Mass Spectrometry

“…Porthidium hyoprora is a representative of this group of venomous snakes; their habitat is the Amazon tropical forest in Brazil, Ecuador, Peru and Colombia [ 3 ]. Proteomic analysis of P. nasutum and P. ophryomegas snake venom and the characterization of toxins from P. lansbergii hutmanni [ 4 , 5 ] have already been performed. However, minimum attention has been paid to the characterization of P. hyoprora venom.…”

The Bushmaster (Cuaima) snake (Lachesis muta muta) of the Neotropical forest: the description of several haemostatic and biological activities in its venom

“…The crude Lmm venom and the CMS-5 fraction were run in electrophoresis under reduced conditions (8 -20 %), using methods [10,12,22] to detect the proteins. For the crude Lmm venom, the polypeptides found were classified into groups, based on their relative molecular masses, establishing that the very high ones were greater than 200 kilodaltons (kDa), high between 200 -66.2 kDa, intermediate 66.2 -21.5 kDa and low between 21.5 -6.5 kDa and very low those below 6 kDa [6].…”