Bmoo FIBMP-I : A New Fibrinogenolytic Metalloproteinase from Bothrops moojeni Snake Venom

Abstract EN

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 (Vmax=0.4596 Uh−1nmol−1±0.1031 and Km=14.59 mg/mL±4.610).