Limited Proteolysis of Fibrinogen by Fibrinogenase from Echis multisquamatis Venom

作者：Chernyshenko V O^*

来源：Protein Journal, 2015, 34(2): 130-134.

DOI：10.1007/s10930-015-9605-2

• Fibrinogen
• Fibrin
• Proteolysis
• Fibrinogenase
• Snake venom
• Limited proteolysis

Previously we purified fibrinogenase from venom of Echis multisquamatis and showed that the enzyme predominantly cleaves B beta Arg42-Ala43 peptide bond of fibrinogen. A much slower hydrolysis of its A alpha-chain was also shown. To evaluate the accessibility of the hydrolysis sites to fibrinogenase's hydrolytic action, the pathway of cleavage of A alpha- and B beta-chains of fibrinogen, monomeric and polymeric fibrin desA and desAB has been investigated using western blot with monoclonal antibodies to B beta 26-42 and A alpha 20-78 of fibrinogen. The data indicated that the B beta Arg42-Ala43 peptide bond is available for cleavage in all forms of fibrin(ogen) with the exception of polymerized fibrin desAB. This is direct evidence of B beta N-domain involvement in formation of protofibrils that makes it inaccessible to protease. The A alpha-chain of fibrinogen remained intact after 3 min of incubation with fibrinogenase. Further incubation resulted in cleaving of the fibrin(ogen) alpha C-regions with the formation of two kinds of degradation products (similar to 30 and similar to 60 kDa). In the case of monomeric fibrin desA or desAB we observed simultaneous hydrolysis of A alpha and B beta-chains and the cleavage of A alpha-chain was more apparent for both forms of polymeric fibrin.

• 出版日期2015-4