Bmoo FIBMP-I: A New Fibrinogenolytic Metalloproteinase fromBothrops moojeniSnake Venom

Abstract

A new fibrinogenolytic metalloproteinase (Bmoo FIBMP-I) was purified from Bothrops moojeni snake venom. This enzyme was isolated through a combination of three chromatographic steps (ion-exchange, molecular exclusion, and affinity chromatography). Analyses by reverse phase chromatography, followed by mass spectrometry, showed the presence of enzyme isoforms with average molecular mass of 22.8 kDa. The SDS-PAGE analyses showed a single chain of 27.6 kDa, in the presence and absence of reducing agent. The protein has a blocked N-terminal. One of the peptides obtained by enzymatic digestion of a reduced and S-alkylated isoform was completely sequenced by mass spectrometry (MS/MS). Bmoo FIBMP-I showed similarity with hemorrhagic factor and several metalloproteinases (MP). This enzyme degraded Aα-chain faster than the Bβ-chain and did not affect the γ-chain of bovine fibrinogen. The absence of proteolytic activity after treatment with EDTA, together with the observed molecular mass, led us to suggest that Bmoo FIBMP-I is a member of the P-I class of the snake venom MP family. Bmoo FIBMP-I showed pH-dependent proteolytic activity on azocasein, but was devoid of coagulant, defibrinating, or hemorrhagic activities. The kinetic parameters of proteolytic activity in azocasein were determined (V max = 0.4596 Uh−1nmol−1 ± 0.1031 and K m = 14.59 mg/mL ± 4.610).

Liquid chromatographic nanofractionation with parallel mass spectrometric detection for the screening of plasmin inhibitors and (metallo)proteinases in snake venoms

“…The fibrin(ogen)olytic effect of snake venom toxins results from different mechanisms, e.g., stimulation of plasminogen activators from endothelial cells [ 7 ], direct plasminogen activation [ 8 , 9 ], or direct cleavage and degradation of fibrinogen and fibrin [ 10 – 13 ]. Most of the molecules which exhibit fibrin(ogen)olytic activities belong to the snake venom serine protease (SVSP) and snake venom metalloproteinase (SVMP) toxin families [ 4 , 14 ].…”

A New Platelet-Aggregation-Inhibiting Factor Isolated fromBothrops moojeniSnake Venom

“…The toxin was isolated using a three-step procedure, including ion-exchange, gel filtration, and affinity chromatographies. Other toxins have been purified using similar procedures, including BmooFIBMP-I [ 30 ] and BmooMP α -1 [ 31 ], both metalloproteinases purified from B. moojeni snake venom. B. moojeni crude venom (200 mg) was applied to ion-exchange chromatography on a DEAE-Sephacel column, which produced eight main protein peaks, designated D1 to D8 ( Figure 1(a) ).…”

Interaction between TNF and BmooMP-Alpha-I, a Zinc Metalloprotease Derived from Bothrops moojeni Snake Venom, Promotes Direct Proteolysis of This Cytokine: Molecular Modeling and Docking at a Glance

“…The peak B1 corresponded to the metalloprotease BmooMP-alpha-I ( Figure 1 C). BmooMP-alpha-I represented a quantity of 8.71% of the whole crude venom of B. moojeni , a significant amount if compared with other fibrin(ogen)olytic enzymes isolated from similar preparations [ 12 , 15 , 16 , 17 ].…”