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Validation of new venom protease assays for translational research (LB589)
Author(s) -
Price Joseph
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.lb589
Subject(s) - envenomation , venom , protease , proteases , metalloproteinase , repurposing , collagenase , elastase , matrix metalloproteinase , pharmacology , antivenom , enzyme , biology , chemistry , microbiology and biotechnology , medicine , biochemistry , ecology
Snake envenomation is an understudied significant world health problem, designated an orphan disease by the WHO. In the U.S. lives are usually saved, but significant tissue is often lost. Overseas without hospitalization and antivenin, lives are forfeit. For most American snake venoms the major pathologies of tissue loss and coagulopathies are due to proteolytic degradation of host tissues and plasma cascades. For HTS leading to therapeutics including drug repurposing, sensitive new substrates may finally allow direct comparisons of venom activities, but only if the reptile enzymes are active upon them, which has been not previously been published. Here I show direct comparisons of representative American venoms strong protease and collagenase, but weak elastase activities using the new technology. Assay CV range 1‐5%. Caseinolytic activity is inhibited by the nonspecific protease inhibitor 1,10‐phenanthroline and by EDTA. So is collagenase activity, consistent with the action of matrix metalloproteases. The Km measured shows that high throughput assays could be done at substrate levels well below the Km for the reaction. Such a system will be invaluable for the testing leading to the repurposing of drugs to treat envenomations. Grant Funding Source : Supported by CHS‐OSU