Purification and characterization of a metalloproteinase, Porthidin-1, from the venom of Lansberg's hog-nosed pitvipers (Porthidium lansbergii hutmanni).
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 LD(50) 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.
Sección de Inmunoquímica, Instituto de Medicina Tropical, Caracas DF 1041, Venezuela.
María E
ME
Girón
Amalid
A
Estrella
Elda E
EE
Sánchez
Jacob
J
Galán
W Andy
WA
Tao
Belsy
B
Guerrero
Ana M
AM
Salazar
Alexis
A
Rodríguez-Acosta
D000595:Amino Acid Sequence; D000818:Animals; D003435:Crotalid Venoms / Q000032:analysis / Q000737:chemistry / Q000201:enzymology* / Q000633:toxicity; D004487:Edema / Q000139:chemically induced / Q000473:pathology; D006470:Hemorrhage / Q000139:chemically induced / Q000473:pathology; D006801:Humans; D007928:Lethal Dose 50; D008297:Male; D045726:Metalloproteases / Q000032:analysis* / Q000737:chemistry / Q000633:toxicity*; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010974:Platelet Aggregation / Q000187:drug effects; D020539:Sequence Analysis, Protein; D021241:Spectrometry, Mass, Electrospray Ionization
D003435:Crotalid Venoms; D045726:Metalloproteases; C558975:porthidin-1, Porthidium lansbergii hutmanni
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