Unlike other sea snakes, Aipysurus eydouxii feeds exclusively on fish eggs. This unusual feeding habit prompted us to search for unique transcripts in their venom glands. In the present study we expressed a novel cysteine-rich secretory protein containing 94 amino acid residues that was identified in its cDNA library. As it induced inflammation and writhing in animals, this protein was named inflamin. It induced two waves of prostanoid production. The first wave peaked at 10 min and 6-oxo PGF1α (prostaglandin F1α) (6-keto PGF1α) was the major product. The second wave, specifically of 6-oxo PGF1α and PGE2 (prostanglandin E2), started after 2 h. In RAW 264.7 cells, COX-1 (cyclo-oxygenase-1) activity showed a transient increase at 10 min and is responsible for the first wave, but its expression was unaffected. COX-2 was induced after 3 h and is responsible for the second wave. Using specific inhibitors, we showed that cPLA2 (calcium-dependent phospholipase A2), and not sPLA2 (secretory phospholipase A2), iPLA2 (calcium-independent phospholipase A2) or DAG (diacylglycerol) lipase, plays a key role in arachidonate release. The cPLA2 activity showed a transient increase of 62% at 10 min; this increase was due to its phosphorylation and not due to an increase in its expression. Thus inflamin, the first member of a new family of snake venom proteins, leads to an increase in the cPLA2 and COX-1 activity resulting in inflammation and pain.
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Research Article|
September 27 2013
Characterization of inflamin, the first member of a new family of snake venom proteins that induces inflammation
Bhaskar Barnwal;
Bhaskar Barnwal
*Department of Biological Sciences, National University of Singapore, Singapore 117543
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R. Manjunatha Kini
R. Manjunatha Kini
1
*Department of Biological Sciences, National University of Singapore, Singapore 117543
†Department of Biochemistry and Molecular Biophysics, Medical College of Virginia, Virginia Commonwealth University, VA 23298, U.S.A.
‡The School of Pharmacy and Medical Sciences, Division of Health Sciences, University of South Australia, Adelaide 5000, Australia
1To whom correspondence should be addressed (email dbskinim@nus.edu.sg).
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Publisher: Portland Press Ltd
Received:
April 29 2013
Revision Received:
July 03 2013
Accepted:
July 08 2013
Accepted Manuscript online:
July 08 2013
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2013 Biochemical Society
2013
Biochem J (2013) 455 (2): 239–250.
Article history
Received:
April 29 2013
Revision Received:
July 03 2013
Accepted:
July 08 2013
Accepted Manuscript online:
July 08 2013
Citation
Bhaskar Barnwal, R. Manjunatha Kini; Characterization of inflamin, the first member of a new family of snake venom proteins that induces inflammation. Biochem J 15 October 2013; 455 (2): 239–250. doi: https://doi.org/10.1042/BJ20130599
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