Functional expression of a novel Kunitz type protease inhibitor from the human blood fluke Schistosoma mansoni

Ranasinghe, Shiwanthi L., Fischer, Katja, Gobert, Geoffrey N. and McManus, Donald P. (2015) Functional expression of a novel Kunitz type protease inhibitor from the human blood fluke Schistosoma mansoni. Parasites and Vectors, 8 408: 1-10. doi:10.1186/s13071-015-1022-z


Author Ranasinghe, Shiwanthi L.
Fischer, Katja
Gobert, Geoffrey N.
McManus, Donald P.
Title Functional expression of a novel Kunitz type protease inhibitor from the human blood fluke Schistosoma mansoni
Formatted title
Functional expression of a novel Kunitz type protease inhibitor from the human blood fluke Schistosoma mansoni
Journal name Parasites and Vectors   Check publisher's open access policy
ISSN 1756-3305
Publication date 2015-08-04
Sub-type Article (original research)
DOI 10.1186/s13071-015-1022-z
Open Access Status DOI
Volume 8
Issue 408
Start page 1
End page 10
Total pages 10
Place of publication London, United Kingdom
Publisher BioMed Central
Language eng
Formatted abstract
Background:  Schistosomes are able to survive for prolonged periods in the blood system, despite continuous contact with coagulatory factors and mediators of the host immune system. Protease inhibitors likely play a critical role in host immune modulation thereby promoting parasite survival in this extremely hostile environment. Even though Kunitz type serine protease inhibitors have been shown to play important physiological functions in a range of organisms these proteins are less well characterised in parasitic helminths.

Methods:  We have cloned one gene sequence from S. mansoni, Smp_147730 (SmKI-1) which is coded for single domain Kunitz type protease inhibitor, E. coli-expressed and purified. Immunolocalisation and western blotting was carried out using affinity purified polyclonal anti-SmKI-1 murine antibodies to determine SmKI-1 expression in the parasite. Protease inhibitor assays and coagulation assays were performed to evaluate the functional roles of SmKI-1.

Results:  SmKI-1 is localised in the tegument of adult worms and the sub-shell region of eggs. Furthermore, this Kunitz protein is secreted into the host in the ES products of the adult worm. Recombinant SmKI-1 inhibited mammalian trypsin, chymotrypsin, neutrophil elastase, FXa and plasma kallikrein with IC50 values of 35 nM, 61 nM, 56 nM, 142 nM and 112 nM, respectively. However, no inhibition was detected for pancreatic elastase or cathepsin G. SmKI-1 (4 μM) delayed blood clot formation, reflected in an approximately three fold increase in activated partial thromboplastin time and prothrombin time.

Conclusions:  We have functionally characterised the first Kunitz type protease inhibitor (SmKI-1) from S. mansoni and show that it has anti-inflammatory and anti-coagulant properties. SmKI-1 is one of a number of putative Kunitz proteins in schistosomes that have presumably evolved as an adaptation to protect these parasites from the defence mechanisms of their mammalian hosts. As such they may represent novel vaccine candidates and/or drug targets for schistosomiasis control.
Keyword Anti-coagulant
Kunitz type protease inhibitor
Schistosoma mansoni
SmKI-1
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
School of Public Health Publications
 
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