title: 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.affiliation: Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar (Mohali), 160062, Punjab, India.
authors.forename: Priyanka
authors.initials: P
authors.lastname: Bajaj
authors.affiliation: Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar (Mohali), 160062, Punjab, India.
authors.forename: Rajan K
authors.initials: RK
authors.lastname: Tripathy
authors.affiliation: Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar (Mohali), 160062, Punjab, India.
authors.forename: Geetika
authors.initials: G
authors.lastname: Aggarwal
authors.affiliation: Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar (Mohali), 160062, Punjab, India.
authors.forename: Ashok K
authors.initials: AK
authors.lastname: Datusalia
authors.affiliation: Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar (Mohali), 160062, Punjab, India.
authors.forename: Shyam S
authors.initials: SS
authors.lastname: Sharma
authors.affiliation: Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar (Mohali), 160062, Punjab, India. apande@niper.ac.in.
authors.forename: Abhay H
authors.initials: AH
authors.lastname: Pande
mesh_terms: D000818:Animals; D043303:Aryldialkylphosphatase / Q000097:blood / Q000737:chemistry / Q000378:metabolism / Q000627:therapeutic use*; D002021:Buffers; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D006868:Hydrolysis; D008297:Male; D051379:Mice; D062025:Organophosphate Poisoning / Q000517:prevention & control*; D010755:Organophosphates / Q000378:metabolism; D058849:Protein Refolding; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000627:therapeutic use*
chemical_list: D002021:Buffers; D010755:Organophosphates; D011994:Recombinant Proteins; D043303:Aryldialkylphosphatase; C468406:PON1 protein, human
keywords: Acetylcholinesterase; Chlorpyrifosoxon; Organophosphate; Paraoxonase 1; Prophylactic