Characterization and structural analysis of a potent anticoagulant phospholipase A2 from Pseudechis australis snake venom

Du, Qianyun Sharon, Trabi, Manuela, Richards, Renee Stirling, Mirtschin, Peter, Madaras, Frank, Nouwens, Amanda, Zhao, Kong-Nan, De Jersey, John, Lavin, Martin F., Guddat, Luke W. and Masci, Paul P. (2016) Characterization and structural analysis of a potent anticoagulant phospholipase A2 from Pseudechis australis snake venom. Toxicon, 111 37-49. doi:10.1016/j.toxicon.2015.12.017


Author Du, Qianyun Sharon
Trabi, Manuela
Richards, Renee Stirling
Mirtschin, Peter
Madaras, Frank
Nouwens, Amanda
Zhao, Kong-Nan
De Jersey, John
Lavin, Martin F.
Guddat, Luke W.
Masci, Paul P.
Title Characterization and structural analysis of a potent anticoagulant phospholipase A2 from Pseudechis australis snake venom
Formatted title
Characterization and structural analysis of a potent anticoagulant phospholipase A2 from Pseudechis australis snake venom
Journal name Toxicon   Check publisher's open access policy
ISSN 1879-3150
0041-0101
Publication date 2016-03-01
Year available 2015
Sub-type Article (original research)
DOI 10.1016/j.toxicon.2015.12.017
Volume 111
Start page 37
End page 49
Total pages 13
Place of publication Kidlington, Oxford, United Kingdom
Publisher Pergamon Press
Collection year 2016
Language eng
Formatted abstract
Pseudechis australis is one of the most venomous and lethal snakes in Australia. Numerous phospholipase A2 (PLA2) isoforms constitute a major portion of its venom, some of which have previously been shown to exhibit not only enzymatic, but also haemolytic, neurotoxic and anticoagulant activities. Here, we have purified a potent anticoagulant PLA2 (identified as PA11) from P. australis venom to investigate its phospholipase, anticoagulant, haemolytic and cytotoxic activities and shown that addition of 11 nM PA11 resulted in a doubling of the clotting time of recalcified whole blood. We have also demonstrated that PA11 has high PLA2 enzymatic activity (10.9 × 104 Units/mg), but low haemolytic activity (0.6% of red blood cells hydrolysed in the presence of 1 nM PA11). PA11 at a concentration lower than 600 nM is not cytotoxic towards human cultured cells. Chemical modification experiments using p-bromophenacyl bromide have provided evidence that the catalytic histidine of PA11 is critical for the anticoagulant activity of this PLA2. PA11 that was subjected to trypsin digestion without previous reduction and alkylation of the disulfide bonds maintained enzymatic and anticoagulant activity, suggesting that proteolysis alone cannot abolish these properties. Consistent with these results, administration of PA11 by gavage in a rabbit stasis thrombosis model increased the clotting time of recalcified citrated whole blood by a factor of four. These data suggest that PA11 has potential to be developed as an anticoagulant in a clinical setting.
Keyword Anticoagulant
Oligomer
Phospholipase A2
Protein purification
Snake venom protein
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

 
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