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A novel coagulation inhibitor from Schistosoma japonicum

Published online by Cambridge University Press:  14 October 2015

SHIWANTHI L. RANASINGHE ,
KATJA FISCHER ,
GEOFFREY N. GOBERT  and
DONALD P. MCMANUS
SHIWANTHI L. RANASINGHE*
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, QLD, Australia School of Public Health, The University of Queensland, Brisbane, QLD, Australia
KATJA FISCHER
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, QLD, Australia
GEOFFREY N. GOBERT
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, QLD, Australia
DONALD P. MCMANUS
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, QLD, Australia
*
*Corresponding Author. Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Locked Bag 2000, Royal Brisbane Hospital, Herston, QLD 4029, Australia. E-mail: shiwanthi.ranasinghe@qimrberghofer.edu.au
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Summary

Little is known about the molecular mechanisms whereby the human blood fluke Schistosoma japonicum is able to survive in the host venous blood system. Protease inhibitors are likely released by the parasite enabling it to avoid attack by host proteolytic enzymes and coagulation factors. Interrogation of the S. japonicum genomic sequence identified a gene, SjKI-1, homologous to that encoding a single domain Kunitz protein (Sjp_0020270) which we expressed in recombinant form in Escherichia coli and purified. SjKI-1 is highly transcribed in adult worms and eggs but its expression was very low in cercariae and schistosomula. In situ immunolocalization with anti-SjKI-1 rabbit antibodies showed the protein was present in eggs trapped in the infected mouse intestinal wall. In functional assays, SjKI-1 inhibited trypsin in the picomolar range and chymotrypsin, neutrophil elastase, FXa and plasma kallikrein in the nanomolar range. Furthermore, SjKI-1, at a concentration of 7·5 µm, prolonged 2-fold activated partial thromboplastin time of human blood coagulation. We also demonstrate that SjKI-1 has the ability to bind Ca++. We present, therefore, characterization of the first Kunitz protein from S. japonicum which we show has an anti-coagulant properties. In addition, its inhibition of neutrophil elastase indicates SjKI-1 have an anti-inflammatory role. Having anti-thrombotic properties, SjKI-1 may point the way towards novel treatment for hemostatic disorders.

Keywords

Kunitz proteintrypsin inhibitoranti-inflammatory agentanti-coagulant
Type
Research Article
Information
Parasitology , Volume 142 , Issue 14 , December 2015 , pp. 1663 - 1672
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Copyright
Copyright © Cambridge University Press 2015 

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