Refolded Recombinant Human Paraoxonase 1 Variant Exhibits Prophylactic Activity Against Organophosphate Poisoning.

Abstract

Organophosphate (OP) compounds are neurotoxic chemicals, and current treatments available for OP-poisoning are considered as unsatisfactory and inadequate. There is an urgent need for the development of more effective treatment(s) for OP-poisoning. Human paraoxonase 1 (h-PON1) is known to hydrolyze a variety of OP-compounds and is a leading candidate for the development of prophylactic and therapeutic agent against OP-poisoning in humans. Non-availability of effective system(s) for the production of recombinant h-PON1 (rh-PON1) makes it hard to produce improved variant(s) of this enzyme and analyze their in vivo efficacy in animal models. Production of recombinant h-PON1 (rh-PON1) using an Escherichia coli expression system is a key to develop variant(s) of h-PON1. Recently, we have developed a procedure to produce active rh-PON1 enzymes by using E. coli expression system. In this study, we have characterized the OP-hydrolyzing properties of refolded rh-PON1(wt) and rh-PON1(H115W;R192K) variant. Our results show that refolded rh-PON1(H115W;R192K) variant exhibit enhanced OP-hydrolyzing activity in in vitro and ex vivo assays and exhibited prophylactic activity in mouse model of OP-poisoning, suggesting that refolded rh-PON1 can be developed as a therapeutic candidate.

• Authors: Priyanka Bajaj, Rajan K Tripathy, Geetika Aggarwal, Ashok K Datusalia, Shyam S Sharma, Abhay H Pande
• Journal: Applied biochemistry and biotechnology (Appl Biochem Biotechnol) Vol. 180 Issue 1 Pg. 165-76 (Sep 2016) ISSN: 1559-0291 [Electronic] United States
• PMID: 27131877 (Publication Type: Journal Article)

Chemical References

• Buffers
• Organophosphates
• Recombinant Proteins
• Aryldialkylphosphatase
• PON1 protein, human

Topics

• Animals
• Aryldialkylphosphatase (blood, chemistry, metabolism, therapeutic use)
• Buffers
• Disease Models, Animal
• Female
• Humans
• Hydrolysis
• Male
• Mice
• Organophosphate Poisoning (prevention & control)
• Organophosphates (metabolism)
• Protein Refolding
• Protein Stability
• Recombinant Proteins (chemistry, metabolism, therapeutic use)