Bmoo FIBMP-I: A New Fibrinogenolytic Metalloproteinase from Bothrops moojeni Snake Venom. (4th November 2012)

• Record Type: Journal Article
• Authors: Torres, F. S.; Rates, B.; Gomes, M. T. R.; Salas, C. E.; Pimenta, A. M. C.; Oliveira, F.; Santoro, M. M.; de Lima, M. E.
• Other Names: Cruz A. Academic Editor.; Ducancel F. Academic Editor.; Utkin Y. N. Academic Editor.
• Is Part Of: ISRN toxicology. Volume 2012(2012)
• Publication Date: 2012-11-04
• Subjects: Toxicology -- Periodicals; Toxicology; Pharmacology; Toxicology; Pharmacology -- Periodicals; Toxicology -- Periodicals; Periodical; Periodicals; Fulltext; Internet Resources; Periodicals; 571.95
• DOI: 10.5402/2012/673941
• Languages: English
• ISSNs: 2090-6188

Abstract

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 − 1 nmol − 1 ± 0.1031 and K m = 14.59 mg/mL ± 4 .610 ).