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

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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.

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Abbreviations

AChE:

Acetylcholinesterase

ATCh:

Acetylthiocholine

CPO:

Chlorpyrifosoxon

Chi-PON1:

Chimeric-paraoxonase 1

CWNA:

Chemical warfare nerve agent

DFP:

Diisopropylflourophosphate

DTNB:

Dithionitrobenzoic acid

h-PON1:

Human paraoxonase 1

rh-PON1:

Recombinant human paraoxonase 1

OP:

Organophosphate

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Acknowledgments

This work was supported by the research grant to AHP from NIPER, S.A.S. Nagar (NPLC-AHP). The authors are grateful to Prof. Dan S. Tawfik (Weizmann Institute of Science, Rehovot, Israel) for kindly providing us with the plasmid containing Chi-PON1 and to Prof. Richard W. James (University Hospital, Geneva, Switzerland) for the gift of monoclonal mouse anti-HuPON1 antibody.

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Authors and Affiliations

  1. Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar (Mohali), 160062, Punjab, India

    Priyanka Bajaj, Rajan K. Tripathy, Geetika Aggarwal & Abhay H. Pande

  2. Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar (Mohali), 160062, Punjab, India

    Ashok K. Datusalia & Shyam S. Sharma

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  1. Priyanka Bajaj
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  3. Geetika Aggarwal
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  6. Abhay H. Pande
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Correspondence to Abhay H. Pande.

Ethics declarations

The experimental protocol was duly approved by Institutional Ethics Committee, NIPER S.A.S. Nagar (IAEC/12/71), and care for animals was carried out according to the guidelines by Committee for the Purpose of Control and Supervision of Experiments on Animals, Government of India.

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The authors declare that they have no conflict of interest.

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Bajaj, P., Tripathy, R.K., Aggarwal, G. et al. Refolded Recombinant Human Paraoxonase 1 Variant Exhibits Prophylactic Activity Against Organophosphate Poisoning. Appl Biochem Biotechnol 180, 165–176 (2016). https://doi.org/10.1007/s12010-016-2091-y

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