An alternative purification method for human serum paraoxonase 1 and its interaction with methidathion

Abstract

In this study, an alternative purification method for human Paraoxonase 1 (hPON1) enzyme was developed using two-step procedures, namely ammonium sulphate precipitation and Sepharose-4B-L-tyrosine-1-aminoanthracene hydrophobic interaction chromatography. SDS-polyacrylamide gel electrophoresis of the enzyme indicates a single band with an apparent MW of 43 kDa. The enzyme was purified 674-fold with a yield of 16%. Furthermore, we examined the in vitro effect of methidathion on the enzyme activity to understand the better inhibitory properties of the compound. Methidathion is a highly toxic insecticide used to control a broad spectrum of agricultural insect and mite pests. IC₅₀ value was found to be 0.130 mM for the pesticide. Methidathion showed a competitive inhibition with Ki of 0.119 mM for paraoxon.

Keywords:

• PON1
• hydrophobic interaction chromatography
• purification
• inhibition
• methidathion

Acknowledgements

This work was supported by Balikesir University Research Project (2013/30).

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article.