Isolation and structural characterization of a new fibrin(ogen)olytic metalloproteinase from Bothrops moojeni snake venom

A proteinase, named BmooMP alpha-I, from the venom of Bothrops moojeni, was purified by DEAE-Sephacel, Sephadex G-75 and heparin-agarose column chromatography. The enzyme was purified to homogeneity as judged by its migration profile in SDS-PAGE stained with coomassie blue, and showed a molecular mass of about 24.5 kDa. Its complete cDNA was obtained by RT-PCR and the 615 bp codified for a mature protein of 205 amino acid residues. The multiple alignment of its deduced amino acid sequence and those of other snake venom metalloproteinases showed a high structural similarly, mainly among class P-IB proteases. The enzyme cleaves the A alpha-chain of fibrinogen first, followed by the B beta-chain, and shows no effects on the gamma-chain. On fibrin, the enzyme hydrolyzed only the beta-chain, leaving the gamma-dimer apparently untouched. It was devoid of phospholipase A(2), hemorrhagic and thrombin-like activities. Like many venom enzymes, it is stable at pH values between 4 and 10 and stable at 70 degrees C for 15 min. The inhibitory effects of EDTA on the fibrinogenolytic activity suggest that BmooMP alpha-I is a metalloproteinase and inhibition by beta-mercaptoethanol revealed the important role of the disulfide bonds in the stabilization of the native structure. Aprotinin and benzamidine, specific serine proteinase inhibitors, had no effect on BmooMP alpha-I activity. Since the BmooMP alpha-I enzyme was found to cause defibrinogenation when administered i.p. on mice, it is expected that it may be of medical interest as a therapeutic agent in the treatment and prevention of arterial thrombosis. (C) 2007 Elsevier Ltd. All rights reserved.

Glazing to entry, Mt. Nebo House

Leadlight windows with stained glass feature panes and timber muntins over french doors with matching leadlight sidelights.

Dining room, Mt. Nebo House

Interior view showing fireplace and mezzanine.

Hallway, Mt. Nebo House

Hallway off main entry to house.

Dormer window, Mt. Nebo House

Arched head leadlight and casement windows with stained glass feature panes and timber muntins. View through window from interior.

Dormer window, Mt. Nebo House

Arched head leadlight window with stained glass feature panes and timber muntins.

Garage-studio interior, Mt. Nebo House

Timber framing and roof skylight detail inside garage-studio.

Upper level, C House, Coorparoo, Brisbane

View along North-West elevation upper level.

Iwan, C House, Coorparoo, Brisbane

As seen from pool deck.

The Nest, C House, Coorparoo, Brisbane

Timber deck with built-in seat overlooking greater landscape beyond.

Pool deck, C House, Coorparoo, Brisbane

View back towards house from deck. with Iwan (right) and filter room (left).

The Nest, C House, Coorparoo, Brisbane

Concrete framework for The Nest (North-West elevation), with timber framework yet to be added.

North-West elevation, C House, Cooparoo, Brisbane

Entry stair in foreground.

Pool deck, C House, Coorparoo, Brisbane

As seen from The Nest above